Compounds for pest control and methods for their use

ABSTRACT

Compounds, compositions, and methods for controlling an arthropod pest population that employ an eremophilane sesquiterpene parent structure are presented. The compounds have minimal adverse or toxic effects on humans, non-human animals, and the natural environment. The compounds may be isolated from natural sources, semi-synthesized from naturally occurring compounds, or completely synthesized. The compounds may be applied directly to a pest, or the locus of a pest, and function as topical or ingestible toxins. Eremophilane sesquiterpenes 13-hydroxy-valencene, valencene-11,12-epoxide, valencene-13-aldehyde, and nootkatone-1,10-11,12-diepoxide are exemplary compounds.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.10/450,024, filed Nov. 10, 2003, now U.S. Pat. No. 7,129,271, which is aU.S. national stage application under 35 U.S.C. §371 of InternationalApplication No. PCT/US01/47736, filed Dec. 7, 2001, which claims thebenefit of U.S. Provisional Patent Application No. 60/254,311, filedDec. 8, 2000, each of which applications is herein incorporated byreference in its entirety.

FIELD

This invention relates to compositions for controlling pest populationsand methods for their use. In addition, this invention relates topesticidal compositions for controlling arthropod pests.

BACKGROUND

Pests such as insects, arachnids, and acarines are detrimental tohumans. Pests include pathogenic organisms that infest mammals andplants, such as those that infest or feed upon plants and livestock,thus causing economic loss or diminishment of plant crops, plantproducts, and livestock. For example, the glassy-winged sharpshooter isa pest that feeds on grape vines, thus diminishing the crop availablefor wine production. Other pests may infest structures such asdwellings, residences, hospitals, and commercial establishments, such asrestaurants and retail stores. These pests may be detrimental to thestructure, such as termites feeding on wooden beams, or simply be anuisance to people who visit or live in infested buildings.Additionally, some pests are vectors for certain diseases that harmhumans and non-human animals, including pets and livestock.

The transmission of vector-born diseases through pests is a problemthroughout the world and is best controlled through the control of thosevectors. For example, the deer tick (Ixodes scapularis) may transmitLyme disease to a host when feeding on the host's blood by passing aninfectious microbe (Borrelia burgdorferi), which lives in the tick'sgut, into the host's bloodstream. A mosquito (Aedes aegypti), prevalentthroughout many tropical and sub-tropical regions of the world, maytransmit Dengue Fever, Yellow Fever, or encephalitis viruses to a hoston which it feeds. The rat flea (Xenopsylla cheopis) is a vector for themicrobe (Yersinia pestis) that causes the Plague.

Pest control is often difficult to achieve. Many pesticides are toxic tohumans and animals and may pollute the environment. Hence, a number ofcommonly used pesticides, such as organophosphates, have been restrictedor made commercially unavailable. Biopesticides derived from naturalsources, such as plants, fungi, or other natural products, offer a saferalternative to chemically synthesized pesticides. Biopesticidesgenerally have fewer health effects and can be better for theenvironment, but many biopesticides offer substantially weaker controlof pests, or control only a limited spectrum of pests, while otherbiopesticides may be environmentally toxic. For example,pyrethrins—pesticides made from the extract of the chyrsanthemumplant—control a wide variety of pests, but are very toxic to fish, suchas bluegill and lake trout. Additionally, pests may become resistant tocertain compounds after continued use; for example, insect resistance topyrethrins already has been observed. Thus, new pest control agentsoffer an alternative for commonly used pesticides.

Therefore, a need exists for an effective pesticide capable ofcontrolling a variety of pests, for example vectors of disease, that isrelatively safe for humans, animals, plants, and the environment.

SUMMARY

Compounds, compositions, and methods for controlling arthropods aredisclosed. The compounds are based on an eremophilane sesquiterpeneparent structure and, when used as pesticides or pest control agents,have minimal adverse or toxic effects on humans, non-human animals, andthe natural environment. The compounds are effective against arthropods,such as insects and acarines, including (but not limited to) members ofthe taxonomic order or subclass Acarina, Diptera, Homoptera, orSiphonoptera.

Certain exemplary pesticidal eremophilane sesquiterpenes described inthis specification are represented by Formulas I, II, III, IV, V and/orVI, as discussed below, and may be isolated from natural sources, suchas Alaska yellow Cedar, Alpinia species, bitter cardamom, and citrusfruits. Additionally, valencene, nootkatol, epinootkatol, nootkatone,and nootkatene may be used as starting materials to synthesize some ofthe described compounds. Thus, the compounds are understood to bebiocides and/or biorational. Particular examples of such pesticidaleremophilane sesquiterpenes are β-hydroxy-valencene,valencene-11,12-epoxide, valencene-13-aldehyde, andnootkatone-1,10-11,12-diepoxide.

Articles of manufacture also are disclosed. In some embodiments, thearticle of manufacture includes a vessel containing a pesticidalcompound, such as a bottle, tube, or can. In other embodiments, thearticle of manufacture includes a device comprising the compound, suchas a flea collar, pest control strip, or rodent trap.

Some embodiments employ a method of controlling an arthropod. In suchembodiments, an arthropod is contacted with a pesticidally effectiveamount of a compound described herein sufficient to cause an adverseeffect on the arthropod. In specific embodiments, the arthropod iskilled or repelled from a locus, though other adverse effects leading topest control, such as inducing sterility or inhibiting oviposition, arepossible.

All compounds described herein may be used in pure form or in the formof a pesticidally acceptable salt or pesticidal composition. Thecompounds may function as pest repellents as well as pesticides, andcertain compounds have a lethal effect on certain arthropods. Thecompounds may be applied directly to a pest, or the locus of a pest, andfunction as topical or ingestible toxins. The compounds may be used tokill pests on, or repel pests from, humans, non-human animals, andplants, including household, industrial, recreational, veterinary,agricultural, silvicultural, horticultural, and environmental uses.Additionally, the compounds may be used to control the spread of diseaseby controlling the arthropod vector for that disease, such as killingthe vector to inhibit transmission of the disease to the host.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B illustrate one method of separating the components ofheartwood essential oil. FIG. 1A illustrates the separation of sevenfractions (I-VII) by a chromatographic process using hexane/diethylether as a solvent, and FIG. 1B illustrates the separation of threefractions by a chromatographic process using CH₂Cl₂/diethyl ether.

DETAILED DESCRIPTION

Compounds, compositions, and methods for controlling an arthropod pestpopulation are provided; The compounds used comprise an eremophilanesesquiterpene and are believed to be substantially nontoxic to bothplants and animals. The pest population may be, for example, apathogenic organism population that feeds upon, damages, irritates, orotherwise adversely affects an animal or plant host. In particularembodiments, the pest functions as a vector for disease. When used aspesticides or pest control agents, these compounds have minimal adverseeffects on humans, domesticated animals, wildlife, and/or the naturalenvironment.

Explanations of Terms

Unless otherwise noted, technical terms are used according toconventional usage. In order to facilitate review of the variousembodiments of the invention, the following explanations of terms areprovided:

As used herein, the singular forms “a,” “an,” and “the,” refer to boththe singular as well as plural, unless the context clearly indicatesotherwise. For example, the term “a pesticidal compound” includes singleor plural pesticidal compounds and can be considered equivalent to thephrase “at least one pesticidal compound.”

As used herein, the term “comprises” means “includes.” For example,“comprising A or B” means “includes A,” “includes B,” or “includes bothA and B.”

The term “alcohol” refers to an aliphatic containing one or morehydroxyl groups, including (but not limited to) ethanol, methanol, orpropanol. A “lower aliphatic alcohol” is an alkane, alkene, or alkyne ofone to six carbon atoms substituted with a hydroxyl group.

The term “aliphatic” refers to straight or branched chain alkanes,alkenes, and alkynes. The term “lower aliphatic” refers to straight orbranched chain alkanes, alkenes, and alkynes of 1 to 10 carbons, forexample 1 to 6 carbon atoms. An aliphatic may be unsubstituted orsubstituted, for example, with an —OH group to form a lower aliphaticalcohol.

The term “alkenyl” refers to a straight or branched chain alkyl radicalcontaining at least two carbon atoms and having one carbon-carbon doublebond. The term “lower alkenyl” refers to an alkenyl containing from twoto six carbon atoms, including (but not limited to): vinyl, 2-propenyl,2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, and 5-hexenyl.

The term “alkoxy” refers to a substituted or unsubstituted alkoxy, wherean alkoxy has the structure —O—R, where R is a substituted orunsubstituted alkyl. In an unsubstituted alkoxy, the R is anunsubstituted alkyl. The term “substituted alkoxy” refers to a grouphaving the structure —O—R, where R is alkyl substituted with anon-interfering substituent. “Lower alkoxy” refers to any alkoxy inwhich R is a lower alkyl. “Thioalkoxy” refers to —S—R, where R issubstituted or unsubstituted alkyl.

The term “alkoxyalkyl” refers to an alkoxy group appended to a loweralkyl radical.

The term “alkyl” refers to a cyclic, branched, or straight chain alkylgroup which, unless otherwise described, contains one to twelve carbonatoms. This term is exemplified by groups such as (but not limited to)methyl, ethyl, n-propyl, isobutyl, t-butyl, pentyl, pivalyl, heptyl,adamantyl, and cyclopentyl. Alkyl groups can be unsubstituted orsubstituted with one or more substituents, for example halogen, alkyl,alkoxy, alkylthio, trifluoromethyl, acyloxy, hydroxy, mercapto, carboxy,aryloxy, aryloxy, aryl, arylalkyl, heteroaryl, amino, alkylamino,dialkylamino, morpholino, piperidino, pyrrolidin-1-yl, piperazin-1-yl,or other functionality. The term “lower alkyl” refers to a cyclic,branched or straight chain alkyl of one to six carbon atoms. This termis further exemplified by such groups as methyl, ethyl, n-propyl,i-propyl, n-butyl, t-butyl, i-butyl (or 2-methylpropyl), sec-butyl,n-pentyl, cyclopropylmethyl, i-amyl, n-amyl, n-pentyl, 1-methylbutyl,2,2-dimethylbutyl, 2-methylpentyl, 2,2-dimethylpropyl, n-hexyl. Loweralkyl groups can be unsubstituted or substituted. One specific exampleof a substituted alkyl is 1,1-dimethyl propyl.

The term “alkylamino” refers to an alkyl group where at least onehydrogen is substituted with an amino group.

The term “amino” refers to a chemical functionality —NR₁R₂ where R₁ andR₂ are independently hydrogen, alkyl, or aryl.

An “analog” is a molecule that differs in chemical structure from aparent compound. Examples include, but are not limited to: a homolog(which differs by an increment in the chemical structure, such as adifference in the length of an alkyl chain); a molecular fragment; astructure that differs by one or more functional groups; or a structurethat differs by a change in ionization, such as a radical. Structuralanalogs are often found using quantitative structure activityrelationships (QSAR), with techniques such as those disclosed inRemington: The Science and Practice of Pharmacology, 19^(th) Edition(1995), chapter 28. A derivative is a biologically active moleculederived from the base molecular structure. A mimetic is a biomoleculethat mimics the activity of another biologically active molecule.Biologically active molecules can include chemical compounds that mimicthe pesticidal activities of the compounds disclosed herein.

An “animal” is a living multicellular vertebrate organism, a categorywhich includes, for example, mammals, reptiles, arthropods, and birds.

The term “aryl” refers to a monovalent unsaturated aromatic carbocyclicgroup having a single ring (e.g., phenyl, benzyl) or multiple condensedrings (e.g., naphthyl or anthryl), which can be unsubstituted orsubstituted with, for example, halogen, alkyl, alkoxy, mercapto (—SH),alkylthio, trifluoromethyl, acyloxy, hydroxy, carboxy, aryloxy, aryl,arylalkyl, heteroaryl, amino, alkylamino, dialkylamino, morpholino,piperidino, pyrrolidin-1-yl, piperazin-1-yl, or other functionality.

Some compounds described herein are pesticides of biological originobtained from a naturally occurring substance or organism. Suchsubstances are commonly called “biocides.” Certain compounds areunderstood to be “biorational,” because the compound is a chemicalsubstance of natural origin that can be synthesized. Pesticides havingan active ingredient selected from compounds according to any ofFormulas I-V that are biorational chemicals qualify for the UnitedStates Environmental Protection Agency's Biorational Program.

“═C” refers to a double-bonded carbon atom.

“Carbonyl-containing” refers to any substituent containing acarbon-oxygen double bond (C═O), including substituents based on —COR or—RCHO where R is an aliphatic or lower aliphatic (such as alkyl or loweralkyl), hydroxyl, or a secondary, tertiary, or quaternary amine.Carbonyl-containing groups include, for example, aldehydes, ketones,carboxylic acids, and esters. Alternatively, “carbonyl-containing group”refers to —R₁COR₂ groups wherein R₁ and R₂ are independently aliphatic,lower aliphatic (such as alkyl or lower alkyl), hydroxyl, or secondary,tertiary, or quaternary amine. Examples include —COOH, CH₂COOH,—CH₂COOCH₃, —CH₂CONH₂, —CH₂CON(CH₃)₂.

“Carboxyl” refers to the radical —COOH, and substituted carboxyl refersto —COR where R is alkyl, lower alkyl, or a carboxylic acid or ester.

“Conjugate” refers to an acid and a base that can convert to each otherby the gain or loss of a proton.

The term “dialkylamino” refers to —N—R—R′ wherein R and R′ areindependently selected from lower alkyl groups.

The term “dialkylaminoalkyl” refers to —N—R—R′, which is appended to alower alkyl radical, wherein R and R′ are independently selected fromlower alkyl groups.

The term “halogen” refers to the elements fluourine, bromine, chlorine,and iodine, and the term “halo” refers to fluoro, bromo, chloro and iodosubstituents.

The term “heterocycle” (or “heterocyclic”) refers to a monovalentsaturated, unsaturated, or aromatic carbocyclic group having a singlering (e.g., benzyl, morpholino, pyridyl or furyl), or multiple condensedrings (e.g., naphthyl, quinolinyl, indolizinyl or benzo[b]thienyl).Additionally, some heterocyles may contain a heteroatom, (such as as N,O, P, or S) in place of a carbon atom within the ring structure. Aheterocycle can be unsubstituted or substituted with, for example,halogen, alkyl, alkoxy, alkylthio, trifluoromethyl, acyloxy, hydroxy,mercapto, carboxy, aryloxy, aryl, arylalkyl, heteroaryl, amino,alkylamino, dialkylamino, morpholino, piperidino, pyrrolidin-1-yl,piperazin-1-yl, or other functionality. Examples include, but are notlimited to, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl,piperazinyl, morpholinyl, thiomorpholinyl, thiazolyl, oxazolyl,isoxazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyridazinyl, andpyrazinyl.

The term “(heterocyclic)alkyl” as used herein refers to a heterocyclicgroup appended to a lower alkyl radical including, but not limited to,pyrrolidinylmethyl and morpholinylmethyl.

The term “host” includes animal, plant, and fungal hosts.

“Hydroxyl” refers to —OH.

“Hydroxyalkyl” refers to —R—OH, wherein R is alkylene, especially loweralkylene (for example in methylene, ethylene, or propylene). Ahydroxyalkyl group may be either linear or branched, such as1-hydroxyisopropyl.

The term “mammal” includes both human and non-human mammals.

The term “═O” indicates a double-bonded oxygen moiety.

“Oxygen-containing group” refers to an R-group containing at least oneoxygen atom. Exemplary, non-limiting oxygen containing groups includeoxygen alone (which may be attached to the molecule by a single ordouble bond), hydroxyl, hydroxylalkyl, or any group containing acarbonyl moiety.

As used herein, the terms “pest,” “pest organism” and “pest population”refer to arthropods, including pathogens and parasites, that negativelyaffect host plants or animals, including humans, by colonizing,attacking, irritating, or feeding upon them, or competing for hostnutrients. The terms “parasite” and “parasitic” refer to all arthropodendoparasites and ectoparasites of hosts. Some pests function as diseasevectors capable of spreading disease to a host population.

Exemplary arthropods include, without limitation, the followingarthropods described according to taxonomic designation and/orvernacular name:

Order Acarina, including Acarus siro, Aceria sheldoni, Aculusschlechtendali, Amblyomma species, Argas species, Boophilus species,Brevipalpus species, Bryobia praetiosa, Calipitrimerus species,Chorioptes species, Dermanyssus gallinae, Eotetranychus carpini,Eriophyes species, Hyalomma species, Ixodes species, Olygonychuspratensis, Ornithodoros species, Panonychus species, Phyllocoptrumoleivora, Polyphagotarsonemus latus, Psoroptes species, Rhipicephalusspecies, Rhizoglyphus species, Sarcoptes species, Tarsonemus species,and Tetranychus species, Dermacentor species.

Order Homoptera, including Aleurothrixus floccosus, Aleyrodes brassicae,Aonidiella species, Aphididae species, Aphis species, Aspidiotusspecies, Bemisia tabaci, Ceroplaster species, Chrysomphalus aonidium,Chrysomphalus dictyospermi, Coccus hesperidum, Empoasca species,Eriosoma lanigerum, Erythroneura spp, Gascardia species, Laodelphaxspecies, Lecanium corni, Lepidosaphes species, Macrosiphus species,Myzus species, Nephotettix species, Nilaparvata species, Paratoriaspecies, Pemphigus species, Planococcus species, Pseudaulacaspisspecies, Pseudococcus species, Psylia species, Pulvinaria aethiopica,Quadraspidiotus species, Rhopalosiphum species, Saissetia species,Scaphoideus species, Schizaphis species, Sitobion species, Trialeurodesvaporariorum, Trioza erytreae, Unaspis citri; and Homalodisca coagulata;

Order Hymenoptera, including Family Formicidae, Family Apidae, andFamily Bombidae, such as Acromyrmex species, Atta species, Cephusspecies, Diprion species, Diprionidae species, Gilpinia polytoma,Hoplocampa species, Lasius species, Monomorium pharaonis, Neodiprionspecies, Solenopsis species, and Vespa species;

Order Diptera, including Family Culicidae, Family Simulidae, FamilyPsychodidae, Family Ceratopogonidae, Family Sarcophagidae, FamilyStreblidae, and Family Nycteribiidae, such as Aedes species, Antherigonasoccata, Bibio hortulanus, Calliphora erythrocephala, Ceratitis species,Chrysomyia species, Culex species, Culex p. pipiens, Cuterebra species,Dacus species, Drosophila species, Fannia species, Gastrophilus species,Glossina species, Hypoderma species, Hyppobosca species, Liriomyzaspecies, Lucilia species, Melanagromyza species, Musca species, Oestrusspecies, Orseolia species, Oscinella frit, Pegomyia hyoscyami, Phorbiaspecies, Rhagoletis pomonella, Sciara species, Stomoxys species, Tabanusspecies, Tannia species, and Tipula species;

Order Siphonaptera, including Ceratophyllus species, Xenopsylla cheopis,Ctenocephalides species, Oropsylla species, Tulex species and Diamanusspecies.

Order Thysanura, including Lepisma saccharina;

Order Lepidoptera; including Acleris species, Adoxophyes species,Aegeria species, Agrotis species, Alabama argulaceae, Amylois species,Anticarsia gemmatalis, Archips species, Argyrotaenia species, Autographaspecies, Busseola fusca, Cadra cautella, Carposina nipponensis, Chilospecies, Choristoneura species, Clysia ambigueua, Cnaphalocrocisspecies, Cnephasia species, Cochylis species, Coleophora species,Crocidolomia binotaus, Cryptophlebia leucotreta, Cydia species, Diatraeaspecies, Diparopsis castanea, Earias species, Ephestia species, Eucosmaspecies, Eupoecilia ambiguena, Euproctis species, Euxoa species,Grapholita species, Hedya nubiferana, Heliothis species, Hellulaandalis, Hyphantria cunea, Keiferia lycopersicella, Leucoptera scitella,Lithocllethis species, Lobesia botrana, Lymantria species, Lyonetiaspecies, Malacosoma species, Mamestra brassicae, Manduca sexta,Operophtera species, Ostrinia nubilalis, Pammene species, Pandemisspecies, Panolis flammea, Pectinophora gossypieua, Phthorimaeaoperculeua, Pieris rapae, Pieris species, Plutella xylostella, Praysspecies, Scirpophaga species, Sesamia species, Sparganothis species,Spodoptera species, Synanthedon species, Thaumetopoea species, Tortrixspecies, Trichoplusia ni, and Yponomeuta species;

Order Coleoptera, including Agriotes species, Anthonomus species,Atomaria linearis, Chaetocnema tibialis, Cosmopolites species, Curculiospecies, Dermestes species, Diabrotica species, Epilachna species,Eremnus species, Leptinotarsa decemlineata, Lissorhoptrus species,Melolontha species, Oryzaephilus species, Otiorhynchus species,Phlyctinus species, Popillia species, Psylliodes species, Rhizoperthaspecies, Scarabeidae, Sitophilus species, Sitotroga species, Tenebriospecies, Tribolium species, and Trogoderma species;

-   -   Order Orthoptera, including Blatta species, Blattella species,        Gryllotalpa species, Leucophaea maderae, Locusta species,        Periplaneta species, and Schistocerca species

Order Isoptera, including Reticulitermes species;

Order Psocoptera, including Liposcelis species;

Order Anoplura, including Haematopinus species, Phthirus pubis;Linognathus species, Pediculus species, Pemphigus species, andPhylloxera species;

Order Mallophaga, including Damalinea species and Trichodectes species;

Order Thysanoptera, including Frankliniella species, Hercinothripsspecies, Taeniothrips species, Thrips palmi, Thrips tabaci andScirtothrips aurantii and

Order Heteroptera, including Cimex species, Distantiella theobroma,Dysdercus species, Euchistus species, Eurygaster species, Leptocorisaspecies, Nezara species, Piesma species, Rhodnius species, Sahlbergellasingularis, Scotinophara species and Triatoma species.

Order Scopriones, including Centruriodes species, Euscorpius species,Parabuthus species, and Vaejovis species.

Order Araneae, including Latrodectus species, Loxosceles species, andBrachypelma species.

Order Hemiptera, including Cimicidae species, Enicocephalidae species,Pentatomidae species, Gerridae species, Saldidae species, Belostomatidaespecies, and Nepidae species.

Class Diplipoda (millipedes).

Class Chilopoda (centipedes).

In particular embodiments, the pest is a member of the taxonomic orderor subclass Acarina, including soft and hard ticks; Diptera, includingTabanidae, anophelines, and culecines; or Siphonoptera. In otherparticular embodiments, the pest belongs to a particular species, suchas Ixodes scapularis (deer tick), Aedes aegypti (mosquito), Xenopsyllacheopis (rat flea), Homalodisca coagulata (glassy-winged sharpshooter),or Culex pithiens (mosquito).

Other exemplary arthropod pests and/or parasites include fleas;mosquitoes; bees, yellow jackets, and wasps; cockroaches, including theAmerican and German cockroach; termites; houseflies and silverleafwhiteflies; lacey-winged sharpshooters or glassy-winged sharpshooters;leaf hoppers, such as the grape or potato leafhoppers; cabbage looper(Lepidoptera); ants, such as the pharaoh ant, argentine ant, carpenterant, and fire ant; stink or lygus bugs; leafminers; western flowerthrips; aphids, such as melon aphids and black bean aphids; arachnids,such as spiders, ticks, and plant mites, including two-spotted spidermites, McDaniel mites, Pacific mites, and European mites.

A “pest control agent” is a compound or composition that controls thebehavior of a pest by causing an adverse effect on that pest, including(but not limited to) physiological damage to the pest; inhibition ormodulation of pest growth; inhibition or modulation of pestreproduction; inhibition or complete deterrence of pest movement into alocus; initiation or promotion of pest movement away from a locus;inhibition or complete suppression of pest feeding activity; or death ofthe pest. A pest control agent may be considered a “pesticide” if itkills at least one individual in a pest population. Additionally, a pestcontrol agent may be non-lethal at a particular concentration or amount(such as a deterrent of pests) and a pesticide at a differentconcentration or amount. A “pesticidally effective amount” of a compoundrefers to an amount that has an adverse biological effect on at leastsome of the pests exposed to the pesticide or pest control agent. Forexample, the effective amount of a compound may be an amount sufficientto repel a pest from a locus, induce sterility in a pest, or inhibitoviposition in a pest. As another example, a “pesticidally effectiveamount” of a compound is capable of killing at least some individuals ina pest population. In specific embodiments, this pesticide is fatal toat least 10% of the pests treated. In particular embodiments, thepesticidally effective amount kills at least 20%, or even 50%, of thepest population. In more particular embodiments, the pesticidallyeffective amount kills over 90%, and nearly 100%, of the pestpopulation. Specific examples of pesticidally effective amounts andtreatments are provided in the Examples below. The term “amountsufficient to inhibit infestation” refers to that amount sufficient todeter, depress, or repel a portion of a pest population so that adisease or infected state in a host population is inhibited or avoided.

A pesticidally effective amount, or an amount sufficient to inhibitinfestation, for a given compound may be determined by routine screeningprocedures employed to evaluate pesticidal activity and efficacy. Somesuch routine screening procedures are discussed in the Examples below orin Maupin, G. O., and Piesman, J., J. Med. Entomol., 31: 319-21 (1994).Particular examples of pesticidal compounds described herein have anLD₅₀ or LC₅₀ of about 65×10⁻³ or less, such as less than 25×10⁻³, lessthan 10×10⁻³, less than 5×10⁻³, less than 5×10⁻³, than 3×10⁻³, or evenless than 1×10⁻³.

Compounds or compositions having a higher level of pesticidal activitycan be used in smaller amounts and concentrations, while compounds orcompositions having a lower level of pesticidal activity may requirelarger amounts or concentrations in order to achieve the same pesticaleffect. Additionally, some compounds or compositions demonstratingpesticidal activity may demonstrate non-lethal pest control effects at adifferent concentration or amount, such as a lower concentration oramount. Non-lethal pest control effects include anti-feeding, reducedfecundity, reduced oviposition, inhibited ecdysis, and sterility.

The term “phenyl” refers to a phenyl group, which may be unsubstitutedor substituted, for example, with a substituent selected from loweralkyl, alkoxy, thioalkoxy, hydroxy and halo.

The term “phenylalkyl” refers to a phenyl group appended to a loweralkyl radical including, but not limited to, benzyl, 4-hydroxybenzyl,4-chlorobenzyl, and 1-naphthylmethyl.

The term “subject” includes both human and veterinary subjects, such asprimates, canines, felines, and rodents.

The term “thioalkoxyalkyl” refers to a thioalkoxy group appended to alower alkyl radical.

Other chemistry terms herein are used according to conventional usage inthe art, as exemplified by The McGraw-Hill Dictionary of Chemical Terms(1985) and The Condensed Chemical Dictionary (1981).

All chemical compounds include both the L- and D-stereoisomers, as wellas either the L- or D-stereoisomer, unless otherwise specified.

Compounds and Compositions

The compounds described herein are terpenes and terpene derivatives,including sesquiterpenes and sesquiterpene derivatives based on a rootstructure having the formula C₁₅H₂₄, though analogs of sesquiterpenesand sesquiterpene derivatives may be produced by additions andsubstitutions of chemical moities. In particular embodiments, thecompounds comprise eremophilane sesquiterpenes, natural product two-ringsesquiterpenes based on eremophilane as a parent structure:

Eremophilane and eremophilane sesquiterpenes are further described in W.M. B. Konst, et al., Flavours (March/April 1975), pages 121-125; andInternational Union of Pure and Applied Chemistry, Nomenclature ofOrganic Chemistry: Section F-Natural Products and Related Compounds,Recommendations 1976, IUPAC Information Bulletin Appendices on TentativeNomenclature, Symbols, Units, and Standards, No. 53, December, 1976(also found in: Eur. J. Biochem. 86: 1-8 (1978)).

The pesticidal eremophilane sesquiterpenes described herein may berepresented by Formula I:

and R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ R₉ R₁₀ R₁₁, and R₁₂ are eachindependently selected from H, ═O, —OH, lower aliphatic, lower aliphaticalcohol, lower aliphatic thiol, carbonyl containing lower aliphatic,thiocarbonyl containing lower aliphatic, lower aliphatic ether, or loweraliphatic epoxide. Additionally, the selection of R₁, R₂, R₃, R₄, R₅,R₆, R₇, R₈ R₉ R₁₀ R₁₁, and R₁₂ should satisfy valence requirements.

In some embodiments, one or two of the bonds C₁-C₂, C₂-C₃, C₃-C₄, C₄-C₅,C₅-C₁₀, C₅-C₆, C₆-C₇, C₇-C₈, C₈-C₉, C₉-C₁₀, or C₁₀-C₁ is a double bond.If C₁₀-C₁ or C₉-C₁₀ is a double bond, then R₅ is absent to satisfyvalence requirements. In some embodiments, one or two ring bonds in theleft ring (i.e., C₁-C₂, C₂-C₃, C₃-C₄, C₄-C₅, C₅-C₁₀, or C₁₀-C₁) is adouble bond, or one or two of the ring bonds in the right ring (i.e.,one of the bonds C₅-C₁₀, C₅-C₆, C₆-C₇, C₇-C₈, C₈-C₉, or C₉-C₁₀) is adouble bond, or one of the ring bonds in each ring is a double bond. Inparticular embodiments, a double bond is located at a particularposition on the ring structure, such as a double bond at either C₁-C₁₀or C₈-C₉ or both C₁-C₁₀ or C₈-C₉. Additionally, any of C₁-C₁₀ may becarbon, CH, or CH₂, as appropriate, to satisfy valence requirements. Inparticular embodiments, C₁₀-C₁ is a double bond, and in more particularembodiments, both C₁-C₁₀ and C₈-C₉ are double bonds.

In some embodiments, a compound according to Formula I is a specificstereoisomer, such as:

In some embodiments, lower aliphatic is a lower alkyl, lower aliphaticalcohol is a lower alkyl alcohol, lower aliphatic thiol is an alkylthiol, carbonyl containing lower aliphatic is a carbonyl containinglower alkyl, thiocarbonyl containing lower aliphatic is a thiocarbonylcontaining lower alkyl, lower aliphatic ether is a lower alkyl ether,and lower aliphatic epoxide is a lower alkyl epoxide.

In some embodiments, one or more of the R-groups on the ring structure(R₃-R₁₁) are H while the remainder are certain substituents, such as ═O,—OH, lower aliphatic alcohol, carbonyl containing lower aliphatic, loweraliphatic ether, or lower aliphatic epoxide. For example, all of R₃-R₁₁may be H, or all but two or three of R₃-R₁₁ may be H. In particularembodiments, R₃, R₄, R₅ R₆, R₈, and R₁₁ are H and R₇, R₉, and R₁₀ areother R-groups, such as ═O, —OH, lower alkyl alcohol, lower alkyl thiol,carbonyl containing lower alkyl, thiocarbonyl containing lower alkyl,lower alkyl ether, or lower alkyl epoxide. In more particularembodiments, R₉, and R₁₀ are lower alkyl, such as methyl. In otherparticular embodiments, R₇ is an oxygen-containing group, such as ═O,—OH, lower aliphatic alcohol, lower alkyl epoxide, or carbonylcontaining lower aliphatic.

In some embodiments, Y is

In particular embodiments, one of the bonds C₁₁-R₁ or C₁-R₂ is a doublebond and R₁₂ is absent. In more particular embodiments, the C₁₁-R₂ bondis a double bond.

In some embodiments, R₁, R₂, and R₁₂ are independently H, ═O, —OH, loweraliphatic (for example, lower alkyl), lower aliphatic alcohol (forexample, lower alkyl alcohol), lower aliphatic ether (for example, loweralkyl ether), carbonyl-containing lower aliphatic (for example,carbonyl-containing alkyl), or lower aliphatic epoxide (for example,lower alkyl epoxide). In particular embodiments, R₂ is ═O, with the bondC₁₁-R₂ being a double bond. In even more particular embodiments, R₁ islower aliphatic alcohol, such as a lower alkyl alcohol (e.g., —CH₂OH),or lower aliphatic, such as lower alkyl (e.g., methyl or ethyl).

In some embodiments, Y is

In particular embodiments, R₁ is H, —OH, lower aliphatic (for example,lower alkyl), lower aliphatic alcohol (for example, lower alkylalcohol), carbonyl-containing lower aliphatic, lower aliphatic ether(for example, lower alkyl ether) or lower aliphatic epoxide (forexample, lower alkyl epoxide). In even more particular embodiments, R₁is lower aliphatic alcohol, such as lower alkyl alcohol (e.g., —CH₂OH)or lower aliphatic, such as lower aklyl (e.g., methyl or ethyl).

Certain exemplary pesticidal eremophilane sesquiterpenes are representedby Formula II:

and, similar to the R-groups of Formula I, R₁, R₂, R₃, R₄, R₅, R₆, R₇,and R₈ are each independently H, ═O, —OH, lower aliphatic, loweraliphatic alcohol, lower aliphatic thiol, carbonyl containing loweraliphatic, thiocarbonyl containing lower aliphatic, lower aliphaticether, or lower aliphatic epoxide. Additionally, the eremophilane ringstructures of compounds described by Formula II may contain double-bondsas described with respect to Formula I.

The compounds of Formula II form a subset of the compounds described inFormula I, and all chemical substitutions and modifications discussed inrelation to Formula I are possible at the corresponding structurepositions on Formula II. For example, the substitutions andmodifications discussed in relation to R₇ of Formula I correspond to R₅of Formula II. As another example, the substitutions and modificationsdiscussed in relation to R₅, R₆, and R₈, of Formula I correspond to R₇of Formula II, and the substitutions and modifications discussed inrelation to, R₁₁, R₃ and R₄ of Formula I correspond to R₆ of Formula II.As yet another example, the substitutions and modifications discussed inrelation to R₁, R₂, and R₁₂ of Formula I correspond to R₁, R₂, and R₈ ofFormula II, respectively, and the substitutions and modificationsdiscussed in relation to R₉, and R₁₀ of Formula I correspond to R₄ andR₈ of Formula II, respectively.

In some embodiments, a compound according to Formula II is a specificstereoisomer, such as:

In some embodiments, lower aliphatic is a lower alkyl; lower aliphaticalcohol is a lower alkyl alcohol; lower aliphatic thiol is a lower alkylthiol; lower aliphatic carboxylic acid is a lower alkyl carboxylic acid;carbonyl containing lower aliphatic is a lower carbonyl containingalkane; thiocarbonyl containing lower aliphatic is a lower thiocarbonylcontaining alkane; lower aliphatic ether is a lower alkane ether; andlower aliphatic epoxide is a lower alkane epoxide.

In some embodiments, R₃, R₄, R₅, R₆, R₇, and R₈, are independently ═O,—OH, lower aliphatic alcohol, carbonyl containing lower aliphatic, loweraliphatic ether, or lower aliphatic epoxide. In alternative embodiments,several of R₃, R₄, R₅, R₆, R₇, and R₈, are substituents and the othersare H. For example, R₃, R₄, and R₅ can be substituents and the others H.In particular embodiments, R₅ is ═O, —OH, lower aliphatic, loweraliphatic alcohol, carbonyl-containing lower aliphatic, lower aliphaticether, or lower aliphatic epoxide. In more particular embodiments, R₅ is═O, or —OH and R₃, and R₄ are lower aliphatic, such as lower alkyl(e.g., methyl or ethyl).

In some embodiments, Y is

In particular embodiments, one of the bonds carbon-R₁ or carbon-R₂ is adouble bond and R₈ is absent. In more particular embodiments, thecarbon-R₂ bond is a double bond and R₈ is absent.

In some embodiments, R₁, R₂, and R₈ are independently H, ═O, —OH, loweraliphatic (for example, lower alkyl), lower aliphatic alcohol (forexample, lower alkyl alcohol), lower aliphatic ether (for example, loweralkyl ether) or lower aliphatic epoxide (for example, lower alkylepoxide). In particular embodiments, R₂ is O and the carbon-R₂ bond is adouble bond. In even more particular embodiments, R₁ is lower aliphaticalcohol, such as a lower alkyl alcohol (e.g., —CH₂OH) or loweraliphatic, such as lower alkyl (e.g., methyl or ethyl).

In some embodiments, Y is

In particular embodiments, R₁ is H, —OH, lower aliphatic (for example,lower alkyl), lower aliphatic alcohol (for example, lower alkylalcohol), lower aliphatic ether (for example, lower alkyl ether) orlower aliphatic epoxide (for example, lower alkyl epoxide). In even moreparticular embodiments, R₁ is lower aliphatic alcohol, such as loweralkyl alcohol (e.g., —CH₂OH) or lower aliphatic, such as lower alkyl(e.g., methyl or ethyl).

In some embodiments, Y is

In particular embodiments, R₁ is independently H, —C═O, —OH, loweraliphatic (for example, lower alkyl), lower aliphatic alcohol (forexample, lower alkyl alcohol), lower aliphatic ether (for example, loweralkyl ether) or lower aliphatic epoxide (for example, lower alkylepoxide); and R₂ is independently O, S, lower aliphatic (for example,lower alkyl), lower aliphatic alcohol (for example, lower alkylalcohol), lower aliphatic ether (for example, lower alkyl ether) orlower aliphatic epoxide (for example, lower alkyl epoxide). Inparticular embodiments, R₂ is O. In even more particular embodiments, R₁is lower aliphatic alcohol, such as a lower alkyl alcohol (e.g., —CH₂OH)or lower aliphatic, such as lower alkyl (e.g., methyl or ethyl).

In some embodiments, R₆ and R₇ form an epoxide group at C₁ and C₁₀ onthe eremophilane ring structure, similar to the joining of R₅ and R₆ atC₁ and C₁₀ described with respect to Formula I. In such embodiments,compounds described by Formula II are based on the structure:

Certain other exemplary pesticidal eremophilane sesquiterpenes arerepresented by Formula III:

and, similar to the R-groups of Formula I, R₁, R₂, R₃, R₄, R₅, R₆, R₇,and R₈ are each independently H, ═O, —OH, lower aliphatic, loweraliphatic alcohol, lower aliphatic thiol, carbonyl containing loweraliphatic, thiocarbonyl containing lower aliphatic, lower aliphaticether, or lower aliphatic epoxide. Additionally, the eremophilane ringstructures of compounds described by Formula III may containdouble-bonds as described with respect to Formulas I and II.

The compounds of Formula III form a subset of the compounds described byFormulas I and II, and all chemical substitutions and modificationsdiscussed in relation to Formulas I and II are possible at thecorresponding structure positions on Formula III.

In some embodiments, a compound according to Formula III is a specificstereoisomer, such as:

In some embodiments, lower aliphatic is a lower alkyl; lower aliphaticalcohol is a lower alkyl alcohol; lower aliphatic thiol is a lower alkylthiol; lower aliphatic carboxylic acid is a lower alkyl carboxylic acid;carbonyl containing lower aliphatic is a lower carbonyl containingalkane; thiocarbonyl containing lower aliphatic is a lower thiocarbonylcontaining alkane; lower aliphatic ether is a lower alkane ether; andlower aliphatic epoxide is a lower alkane epoxide.

In some embodiments, R₃, R₄, R₅, R₆, R₇, and R₈, are independently ═O,—OH, lower aliphatic alcohol, carbonyl containing lower aliphatic, loweraliphatic ether, or lower aliphatic epoxide. In alternative embodiments,several of R₃, R₄, R₅, R₆, R₇, and R₈, are substituents and the othersare H. For example, R₃, R₄, and R₅ can be substituents and the others H.In particular embodiments, R₅ is ═O, —OH, lower aliphatic, loweraliphatic alcohol, carbonyl-containing lower aliphatic, lower aliphaticether, or lower aliphatic epoxide. In more particular embodiments, R₅ is═O, or —OH and R₃, and R₄ are lower aliphatic, such as lower alkyl(e.g., methyl or ethyl).

In some embodiments, Y is

In particular embodiments, one of the bonds carbon-R₁ or carbon-R₂ is adouble bond and R₈ is absent. In more particular embodiments, thecarbon-R₂ bond is a double bond and R₈ is absent.

In some embodiments, R₁, R₂, and R₈ are independently H, ═O, —OH, loweraliphatic (for example, lower alkyl), lower aliphatic alcohol (forexample, lower alkyl alcohol), lower aliphatic ether (for example, loweralkyl ether) or lower aliphatic epoxide (for example, lower alkylepoxide). In particular embodiments, R₂ is O and the carbon-R₂ bond is adouble bond. In even more particular embodiments, R₁ is lower aliphaticalcohol, such as a lower alkyl alcohol (e.g., —CH₂OH) or loweraliphatic, such as lower alkyl (e.g., methyl or ethyl).

In some embodiments, Y is

In particular embodiments, R₁ is H, —OH, lower aliphatic (for example,lower alkyl), lower aliphatic alcohol (for example, lower alkylalcohol), lower aliphatic ether (for example, lower alkyl ether) orlower aliphatic epoxide (for example, lower alkyl epoxide). In even moreparticular embodiments, R₁ is lower aliphatic alcohol, such as loweralkyl alcohol (e.g., —CH₂OH) or lower aliphatic, such as lower alkyl(e.g., methyl or ethyl).

In some embodiments, Y is

In particular embodiments, R₁ is independently H, —C═O, —OH, loweraliphatic (for example, lower alkyl), lower aliphatic alcohol (forexample, lower alkyl alcohol), lower aliphatic ether (for example, loweralkyl ether) or lower aliphatic epoxide (for example, lower alkylepoxide); and R₂ is independently O, S, lower aliphatic (for example,lower alkyl), lower aliphatic alcohol (for example, lower alkylalcohol), lower aliphatic ether (for example, lower alkyl ether) orlower aliphatic epoxide (for example, lower alkyl epoxide). Inparticular embodiments, R₂ is O. In even more particular embodiments, R₁is lower aliphatic alcohol, such as a lower alkyl alcohol (e.g., —CH₂OH)or lower aliphatic, such as lower alkyl (e.g., methyl or ethyl).

Certain other exemplary pesticidal eremophilane sesquiterpenes arerepresented by Formula IV:

and, similar to the R-groups of Formula I, R₁, R₂, R₃, R₄, R₅, and R₆are each independently H, ═O, —OH, lower aliphatic, lower aliphaticalcohol, lower aliphatic thiol, carbonyl-containing lower aliphatic,thiocarbonyl-containing lower aliphatic, lower aliphatic ether, or loweraliphatic epoxide.

The compounds described by Formula IV form a subset of the compoundsdescribed by Formulas I and II. All chemical substitutions andmodifications discussed in relation to Formulas I and II are possible atthe corresponding positions on Formula IV.

In some embodiments, a compound according to Formula IV is a particularstereoisomer, such as:

Other exemplary pesticidal eremophilane sesquiterpenes are representedby Formula V:

R₁ is lower alkyl, lower alkyl alcohol, or carbonyl-containing loweralkyl; R₂ is lower alkyl, or O; R₃ is lower alkyl; R₄ is lower alkyl;and R₅ is H, —OH, ═O; lower alkyl alcohol, or carbonyl-containing loweralkyl; or Y is

R₁ is lower alkyl, lower alkyl alcohol, or carbonyl-containing loweralkyl; R₃ is lower alkyl; R₄ is lower alkyl; and R₅ is H, —OH, ═O; loweralkyl alcohol, or carbonyl-containing lower alkyl; or Y is

R₁ is H, ═O, —OH, lower alkyl, lower alkyl alcohol, lower alkyl ether,lower alkyl aldehyde, lower alkyl ketone, or lower alkyl epoxide; R₂ isH, ═O, —OH, lower alkyl, lower alkyl alcohol, lower alkyl ether, loweralkyl aldehyde, lower alkyl ketone, or lower alkyl epoxide; R₃ is loweralkyl; R₄ is lower alkyl; and R₅ is H, —OH, ═O; lower alkyl alcohol,carbonyl-containing lower alkyl; R₈ is H, —OH, lower aliphatic, or loweraliphatic alcohol. However, if either the carbon-R₁ or carbon-R₂ is adouble bond, then R₈ is absent. For example, in some embodiments, thecarbon-R₂ bond is a double bond, such as embodiments where R₂ is O, thecarbon-R₂ bond is a double bond, and R₁ is a lower alkyl alcohol (e.g.,—CH₂OH) or lower alkyl (e.g., methyl or ethyl).

The compounds described by Formula V form a subset of the compoundsdescribed by Formulas I and II, and all chemical substitutions andmodifications discussed in relation to Formulas I and II are possible atthe corresponding structure positions on Formula V.

In some embodiments, a compound according to Formula V is a particularstereoisomer, such as:

In some embodiments, R₃ is lower alkyl, such as methyl. In someexamples, R₄ is lower alkyl (such as methyl) or lower alkyl alcohol(such as —CH₂OH). In some examples, R₅ is H. In other embodiments, R₅ isH or —OH, Y is

and R₁ is lower alkyl, such as methyl or ethyl, or lower alkyl alcohol,such as ethyl alcohol.

In other embodiments, Y is

and R₁ is lower alkyl alcohol or lower alkyl alcohol. In suchembodiments, R₃ and R₄ may independently be lower alkyl, and R₅ may beH.

In other embodiments, Y is

R₁ is lower alkyl alcohol and R₂ is O or lower alkyl. In suchembodiments, R₃ and R₄ may independently be methyl, and R₅ may be H.

Still other exemplary pesticidal eremophilane sesquiterpenes arerepresented by Formula VI:

and, similar to the R-groups of Formula I, R₁, R₂, R₃, R₄, R₅, and R₈are each independently H, ═O, —OH, lower aliphatic, lower aliphaticalcohol, lower aliphatic thiol, carbonyl-containing lower aliphatic,thiocarbonyl-containing lower aliphatic, lower aliphatic ether, or loweraliphatic epoxide.

The compounds described by Formula VI form a subset of the compoundsdescribed by Formulas I and II, and all chemical substitutions andmodifications discussed in relation to Formulas I and II are possible atthe corresponding structure positions on Formula VI.

In some embodiments, a compound according to Formula VI is a particularstereoisomer, such as:

In particular embodiments, Y is

and R₁ is lower alkyl or lower alkyl alcohol; R₂ is lower alkyl, or O;R₃ is lower alkyl; R₄ is lower alkyl; and R₅ is H, —OH, or ═O.

In other particular embodiments, Y is

R₁ is lower alkyl or lower alkyl alcohol; R₃ is lower alkyl; R₄ is loweralkyl; and R₅ is H, —OH, or ═O.

In still other particular embodiments, Y is

R₁ is H, ═O, —OH, lower alkyl, lower alkyl alcohol, lower alkyl ether,or lower alkyl epoxide; R₂ is H, ═O, —OH, lower alkyl, lower alkylalcohol, lower alkyl ether, or lower alkyl epoxide; R₃ is lower alkyl;R₄ is lower alkyl; R₅ is H, —OH, or ═O; and R₈ is H, —OH, lower alkyl,or lower alkyl alcohol. However, if either the carbon-R₁ or carbon-R₂ isa double bond, then R₈ is absent. For example, in some embodiments, thecarbon-R₂ bond is a double bond. In even more particular embodiments, R₂is O, the carbon-R₂ bond is a double bond, and R₁ is a lower alkylalcohol (e.g., —CH₂OH) or lower alkyl (e.g., methyl or ethyl).

While some of the compounds encompassed by Formulas I-VI are known—forexample, valencene, nootkatol, epinootkatol, and nootkatone—many othercompounds are novel. Examples of novel compounds include, but are notlimited to, 13-hydroxy-valencene, valencene-11,12-epoxide,valencene-13-aldehyde, and nootkatone-1,10-11,12-diepoxide.

Valencene, nootkatol, epinootkatol, nootkatone, and nootkatene arecommercially available and may be isolated from natural sources. Forexample, nootkatone may be prepared according to the methods andprocesses of U.S. Pat. No. 5,847,226 and WO 97/22575A1, and valenceneand nootkatone may be obtained from Bedoukian Research Inc., of Danbury,Conn. These compounds are known to be nontoxic to humans and non-humananimals. For example, nootkatone is used as a fragrance and foodflavoring.

The compounds described by Formulas I, II, III, IV, V and/or VI,including 13-hydroxy-valencene, valencene-11,12-epoxide,valencene-13-aldehyde, and nootkatone-1,10-11,12-diepoxide, may beisolated from natural sources, such as Alaska yellow Cedar, Alpiniaspecies, bitter cardamom, and citrus fruits (e.g., grapefruit), may besemi-synthesized from compounds isolated from such natural sources, ormay be completely synthesized. Example 1 provides one method forisolating compounds from Alaska yellow cedar, and Examples 11-14illustrate semi-synthesis of the compounds.

Compounds described herein may be described by their common names,numerical compound identifiers, or IUPAC names. Certain, non-limitingexemplary compounds are listed in Table 1.

TABLE 1 Common Name Number IUPAC Name valencene compound 54βH,5α-eremophlia-1(10),11- diene nootkatene compound 64βH,5α-eremophlia-1,9,11- triene nootkatone compound 74βH,5α-eremophlia-1(10),11- dien-2-one 13-hydroxy-valencene compound 104βH,5α-eremophlia-1(10)-ene nootkatol compound 12 2α-hydroxy-4βH,5α-eremophlia-1(10),11-dien valencene-11,12- compound 1311,12-epoxy-4βH,5α- epoxide eremophlia-1(10)-ene valencene-13-aldehydecompound 15 4βH,5α-eremophlia-1(10),11- diene-13-ol nootkatone-11,12-compound 16 11,12-epoxy-4βH,5α- epoxide eremophlia-1(10)-en-2-onenootkatone-1,10- compound 17 1,10-epoxy-4βH,5α-eremophlia- epoxide11-en-2-one nootkatone-1,10- compound 18 1,10-(11,12)-diepoxy-4βH,5α-11,12-diepoxide eremophlia-2-one

Compounds 10, 13, 15, and 18 (13-hydroxy-valencene,valencene-11,12-epoxide, valencene-13-aldehyde, andnootkatone-1,10-11,12-diepoxide) are understood to be novel compounds.

In some embodiments, the addition of oxygen-containing groups increasesthe bioactivity of the compound. Exemplary oxygen-containing groupsinclude double-bond oxygen moities and hydroxy-containing orcarbonyl-containing groups, such as ═O, —OH, lower aliphatic alcohol(such as methyl alcohol or ethyl alcohol), lower aliphatic carboxylicacid, carbonyl-containing lower aliphatic (such as a ketone oraldehyde), lower aliphatic ether, or lower aliphatic epoxide. In otherembodiments, the addition of R-groups containing hydrogen-bonding atomsor functional groups, including both hydrogen bond donors and hydrogenbond acceptors, increases the bioactivity of the compound. It isunderstood that some R-groups may be both oxygen-containing groups andhydrogen-bond donors or acceptors. Additionally, sulfur-containing groupanalogous to the oxygen-containing groups (where the group contains asulfur atom in the position otherwise occupied by an oxygen atom)described herein increase the bioactivity of compounds in someembodiments.

Tables 2 and 3 present particular embodiments of compounds according toFormula I:

In the exemplary compounds listed in Table 2, the C₁-C₁₀ bond is adouble bond and R₅ is absent. Additionally, some of these compounds alsoare encompassed and may be described by Formula II, III, IV, V and/orVI.

TABLE 2 Compound 51 Compound 52 Compound 53 Compound 54 Compound 55 R₁CH₂OH CH₂OH CH₂OH CH₂OH CH₃ R₂ CH₃ CH₃ CH₂OH CH₂OH CH₃ R₃ H H H H H R₄ HH H H H R₆ H H H H H R₇ ═O OH H ═O OH R₈ H H H H H R₉ CH₃ CH₃ CH₃ CH₃CH═O R₁₀ CH₃ CH₂OH CH═O CH₂OH CH₃ R₁₁ H H H H H R₁₂ OH H H OH OHCompound 56 Compound 57 Compound 58 Compound 59 Compound 60 R₁ CH₂OH CH₃CHOCH₃ CH₂CH═CH₃ CH₂CH₂COOH R₂ CHO COCH₃ CH₃ CH₂COOH CH₂OH R₃ H H H H HR₄ H H H H H R₆ H H H H H R₇ ═O OH H OH H R₈ H H H H H R₉ CH₂OH CH═OCH═O CH₂OH CH₃ R₁₀ CH₃ CH═O CH₃ CH₂OH CH═O R₁₁ H H H H H R₁₂ H OHCH₂CH₂OH CH₂COOH H

TABLE 3

R₁ is CH₂OH, CHO, lower aliphatic epoxide, or lower alipahtic ether. R₂is CH₂. R₆ is CH₂OH, CHO, or lower aliphatic ether.

R₁ is ═O, CHOH, C═O, or lower alipahtic ether. R₂ is —OH, CH₂OH, CHO, orcarbonyl-containing lower alipahtic.

R₁ is CHOH, lower aliphatic, or carbonyl-containing lower aliphatic. R₂is CH₃ or other lower alkyl, —OH, or carbonyl- containing lower alkyl.

R₁ is lower aliphatic alcohol, lower aliphatic thiol,carbonyl-containing lower aliphatic, or thiocarbonyl- containing loweraliphatic. R₂ is H, —OH, CH₂OH, or ═O.

R₁ is CH₃, CH₂OH, CH═CHOH, COOH. R₂ is CH, OH, or ═O.

As illustrated by the exemplary compounds of Tables 1-3, particularembodiments employ compounds where R₁ and R₂ are independently loweralkyl, lower alcohol, or lower alkenyl; R₃, R₄, and R₆ are H; R₇ is H,—OH, or ═O; R₈ is H; R₉ and R₁₀ are independently lower alkyl, loweralcohol, or lower aldehyde; R₁₁ is H; and R₁₂ is H, —OH, lower alcohol,or carbonyl-containing lower alkyl.

Specific examples of compounds encompassed by Formulas I-VI includethose listed in Table 4, though this list of compounds is merelyrepresentative and not exhaustive.

TABLE 4

13-hydroxy-valencene Compound 10

valencene-11,12-epoxide Compound 13

valencene-13-aldehyde Compound 15

nootkatone-11,12-epoxide Compound 16

nootkatone-1,10-epoxide Compound 17

nootkatone-1,10-11,12-diepoxide Compound 18Pesticidally Acceptable Salts and Compositions

All compounds described herein may be used in pure form or in the formof a pesticidally acceptable salt. Pesticidally acceptable salts of thecompound of any of Formulas I-VI may be salts of organic or inorganicacids, such as hydrochloric acid, hydrobromic acid, sulfuric acid,nitric acid, perchloric acid, phosphoric acid, formic acid, acetic acid,trifluoroacetic acid, oxalic acid, malonic acid, toluenesulfonic acid,benzoic acid, terpenoid acids (e.g., abiotic acid), or natural phenolicacids (e.g., gallic acid and its derivatives). Additionally, thecompound of any of Formulas I-VI may be included as an active ingredientwithin a composition, for example, a pesticide or pest control agent, ina free form or in the form of a a pesticidally acceptable salt.

A pesticidal composition includes one or more of the above-describedcompounds and a pesticidally acceptable carrier, additive, or adjuvant,and the pesticidal composition may function as a pesticide or pestcontrol agent.

Such pesticidal compositions may be in the form of a solid, liquid, gas,or gel. If a solid composition is created, suitable solid carriersinclude agriculturally useful and commercially available powders. Liquidcompositions may be aqueous or non-aqueous, depending on the needs ofthe user applying the pesticidal composition, and liquids may exist asemulsions, suspensions, or solutions. Exemplary compositions include(but are not limited to) powders, dusts, granulates, topical oils,encapsulations, emulsifiable concentrates, suspension concentrates,directly sprayable or dilutable solutions, coatable pastes, diluteemulsions, wettable powders, soluble powders, dispersible powders, orfumigants.

The particle or droplet size of a particular composition may be alteredaccording to its intended use. The pesticidal composition also mayinclude an apparatus for containing or dispersing the compound orcomposition, such as a storage kit, fumigant bottle (such as thecommonly named “flea bomb”), or insect trap.

Pesticidally acceptable carriers, additives, and adjuvants includestabilizers, preservatives, antioxidants, extenders, solvents,surfactants, antifoaming agents, viscosity regulators, binders, tackers,or other chemical agents, such as fertilizers, antibiotics, fungicides,nematicides, or herbicides. Such carriers, additives, and adjuvants maybe used in solid, liquid, gas, or gel form, depending on the embodimentand its intended application. Pesticidally acceptable adjuvants arethose materials that assist or enhance the action of a compound orcomposition. Surfactants and antifoaming agents are just two examples ofpesticidally acceptable adjuvants. However, any particular material mayalternatively function as a “carrier,” “additive,” or “adjuvant” inalternative embodiments, or may fulfill more than one function.

Certain additives, carriers, or adjuvants may be active or inactivematerials or substances. In some instances, the efficacy of acomposition may be increased by adding one or more other components thatminimize toxicity to hosts or increase the anti-pest effect of thecomposition.

Additionally, the composition may include plural pesticidal compounds.Such a composition includes a compound as described herein and a secondpesticidal compound, and the second pesticidal compound also may be acompound as described herein, or may be any other type or class ofpesticide (e.g., an organophosphate or pyrethrin).

In certain compositions, the second pesticidal compound, additive,carrier, or adjuvant provides a synergistic effect by increasing theefficacy of the pesticidal composition more than the additive amount. Asjust one, non-limiting example, a composition containing both 1%nootkatone and 1% 13-hydroxy-valencene by weight that is more than twiceas effective—such as four times as effective or ten times aseffective—than a composition containing only 1% nootkatone or 1%13-hydroxy-valencene by weight demonstrates a synergistic effect.

The following list of exemplary carriers, additives, and adjuvants ismeant to be illustrative, not exhaustive.

Suitable solid carriers, such as those used for dusts and dispersiblepowders, include natural mineral fillers such as calcite, talcum,kaolin, montmorillonite, and attapulgite. Highly dispersed silicic acidsor highly dispersed absorbent polymers may be added to such carriers.Granulated materials of inorganic or organic nature may be used, such asdolomite or pulverized plant residues. Suitable porous granulatedadsorptive carriers include pumice, broken brick, sepiolite, andbentonite. Additionally, nonsorbent carriers, such as sand, may be used.Some solid carriers are biodegradable polymers, including biodegradablepolymers that are digestible or degrade inside an animal's body overtime.

Suitable liquid carriers, such as solvents, may be organic or inorganic.Water is one example of an inorganic liquid carrier. Organic liquidcarriers include vegetable oils and epoxidized vegetable oils, such asrape seed oil, castor oil, coconut oil, soybean oil and epoxidized rapeseed oil, castor oil, coconut oil, soybean oil, and other essentialoils. Other organic liquid carriers include silicone oils, aromatichydrocarbons, and partially hydrogenated aromatic hydrocarbons, such asalkylbenzenes containing 8 to 12 carbon atoms, including xylenemixtures, alkylated naphthalenes, or tetrahydronaphthalene. Aliphatic orcycloaliphatic hydrocarbons, such as paraffins or cyclohexane, andalcohols, such as ethanol, propanol or butanol, also are suitableorganic carriers. Gums, resins, and rosins used in forest productsapplications and naval stores (and their derivatives) also may be used.Additionally, glycols, including ethers and esters, such as propyleneglycol, dipropylene glycol ether, diethylene glycol, 2-methoxyethanol,and 2-ethoxyethanol, and ketones, such as cyclohexanone, isophorone, anddiacetone alcohol may be used. Strongly polar organic solvents includeN-methylpyrrolid-2-one, dimethyl sulfoxide, and N,N-dimethylformamide.

Suitable surfactants may be nonionic, cationic, or anionic, depending onthe nature of the compound used as an active ingredient. Surfactants maybe mixed together in some embodiments. Nonionic surfactants includepolyglycol ether derivatives of aliphatic or cycloaliphatic alcohols,saturated or unsaturated fatty acids and alkylphenols. Fatty acid estersof polyoxyethylene sorbitan, such as polyoxyethylene sorbitan trioleate,also are suitable nonionic surfactants. Other suitable nonionicsurfactants include water-soluble polyadducts of polyethylene oxide withpolypropylene glycol, ethylenediaminopolypropylene glycol andalkylpolypropylene glycol. Particular nonionic surfactants includenonylphenol polyethoxyethanols, polyethoxylated castor oil, polyadductsof polypropylene and polyethylene oxide, tributylphenol polyethoxylate,polyethylene glycol and octylphenol polyethoxylate. Cationic surfactantsinclude quaternary ammonium salts carrying, as N-substituents, an 8 to22 carbon straight or branched chain alkyl radical. The quaternaryammonium salts carrying may include additional substituents, such asunsubstituted or halogenated lower alkyl, benzyl, or hydroxy-lower alkylradicals. Some such salts exist in the form of halides, methyl sulfates,and ethyl sulfates. Particular salts include stearyldimethylammoniumchloride and benzyl bis(2-chloroethyl)ethylammonium bromide. Suitableanionic surfactants may be water-soluble soaps as well as water-solublesynthetic surface-active compounds. Suitable soaps include alkali metalsalts, alkaline earth metal salts, and unsubstituted or substitutedammonium salts of higher fatty acids. Particular soaps include thesodium or potassium salts of oleic or stearic acid, or of natural fattyacid mixtures. Synthetic anionic surfactants include fatty sulfonates,fatty sulfates, sulfonated benzimidazole derivatives, andalkylarylsulfonates. Particular synthetic anionic surfactants includethe sodium or calcium salt of ligninsulfonic acid, of dodecyl sulfate,or of a mixture of fatty alcohol sulfates obtained from natural fattyacids. Additional examples include alkylarylsulfonates, such as sodiumor calcium salts of dodecylbenzenesulfonic acid, ordibutylnaphthalenesulfonic acid. Corresponding phosphates for suchanionic surfactants are also suitable.

The concentration of a compound, such as a compound according to any ofFormulas I-VI, which serves as an active ingredient, may vary accordingto particular compositions and applications. In a number of embodiments,the percentage by weight of the active ingredient will be from about0.1% to about 90%. A suitable amount for a particular application may bedetermined using bioassays for the particular pest intended to becontrolled. Higher concentrations are usually employed for commercialpurposes or products during manufacture, shipment, or storage; suchembodiments have concentrations at least about 10%, or from about 25% toabout 90% by weight. Prior to use, a highly concentrated formulation maybe diluted to a concentration appropriate for the intended use, such asfrom about 0.1% to 10%, or from about 1% to 5%. In any such formulation,the active ingredient may be a compound according to any of FormulasI-VI, a corresponding pesticidally acceptable salt, or a mixturethereof.

The compounds have deterrent, repellent, and/or toxic effects on certainpest targets and may function as pest repellents or pest control agents,as well as pesticides. Certain compounds have a lethal effect onspecific pests. Unlike a number of commercially available pesticides,many compositions have an active ingredients (such as a compoundaccording to Formula I, II, III, IV, V and/or VI) that are substantiallynontoxic to humans and domesticated animals and that have minimaladverse effects on wildlife and the environment.

The efficacy of a subject compound or composition is determined from anadverse effect on the pest population, including (but not limited to)physiological damage to a pest, inhibition or modulation of pest growth,inhibition or modulation of pest reproduction by slowing or arrestingproliferation, inhibition or complete deterrence of pest movement into alocus, initiation or promotion of pest movement away from a locus,inhibition or elimination of pest feeding activity, or death of thepest, all of which are encompassed by the term “controlling.” Thus, acompound or composition that controls a pest (i.e., a pest control agentor pesticide) adversely affects its presence, status, and/orphysiological condition at a locus. The efficacy and quantity of apesticidally effective amount for a given compound may be determined byroutine screening procedures employed to evaluate pesticidal activityand efficacy, such as those screening described in the Examples.

Efficacy and appropriateness of a compound also may be assessed bytreating an animal, plant, or environmental locus with a compound orcomposition described herein and observing the effects on the infestingpest population and any harm to plants or animals contacted by thecompound, such as phytotoxicity to plants, toxicity to animals, ordermal sensitivity to animals. For example, in certain embodiments,compounds or compositions are directly applied to a host plant or animalactually or potentially infested with a pest. In such embodiments, theefficacy of the compound or composition may be monitored by examiningthe state of host or environmental locus infestation by the pestpopulation before and after application in light of physiological damageto an animal or plant host infected by the pest population found withinthe environmental locus. Additionally, the appropriateness of a compoundor composition may be assessed by observing any adverse effects to theperson applying the composition to an infested plant, animal, orenvironmental locus. In particular embodiments, the effective amount ofa compound or composition meets the mortality, modulation, or controlcriteria above, and has minimal or no adverse effect on plants,non-human animals, or humans that may come into contact with thecompound or composition.

The compounds and compositions have a broad range of biocidal effects,such as pesticidal activity against one or more pests, and certaincompounds and/or compositions may be more effective on some pests thanothers. Some compounds according to any of Formulas I-VI, orcompositions containing such compounds, may be partially or totallyineffective against some pests at certain concentrations. However, anydifferences in efficacy should not in any way detract from the utilityof these compounds or compositions, or their methods of use, since someof these compounds or compositions may function as broad, general actingpesticides, while other compounds or compositions may function asspecific or selective pesticides. The Examples set forth belowillustrate methods by which the degree of selectivity of pesticidalactivity may be readily ascertained.

The subject compounds and compositions offer several advantages overcurrently used pesticides. These naturally-occurring compounds may beisolated from a variety of plant sources, including Alaska yellow cedarand grapefruit, and generally exhibit a very high LD₅₀ againstnon-arthopod animals. Thus, these compounds are relatively nontoxic tohumans, domesticated animals and livestock, birds, fish, and otherwildlife.

The compounds and compositions described herein may be used to controlor eliminate crop pests (and may be used up to harvest), to control thegrowth of pests on harvested crops and stored foods, and for controllingpests in natural and artificial environments. The compound orcomposition may be applied to plant and animal parts (e.g., skin, fur,feathers, scales, leaves, flowers, branches, fruits) and to objectswithin an environment that come into contact with a pest. Additionally,the compound or composition may be included as part of an object held orplaced upon a prospective host plant or animal to inhibit pestinfestation, such as a collar, clothing, or supporting mechanism (e.g.,a stake supporting a seedling tree, a rose trellis, or a cage forsupporting a tomato plant).

The compounds and compositions have useful inhibitory and/or curativeproperties in the field of pest control, even at low concentrations, andmay be used as part of an integrated pest management (IPM) program.These and other methods of using the compounds and compositions arefurther described below.

The compounds and compositions function as topical or ingestible toxinseffective against all developmental stages of arthropod pests, such asinsects and acarines (i.e., members of taxonomic orders Insecta andAcarina). The onset of the pesticidal action of the compounds andcompositions may follow directly (e.g., kill a pest within a shortamount of time) or onset of pesticidal action may occur some time afterthe pest has initially contacted the compound or composition.

Methods of Use

The compounds and compositions according to any of Formulas I-VI may beused as pesticides, including acaricides and insecticides, or may beused as agents to control pests, such as pest repellents. Someembodiments of using these compounds and compositions cover a range ofapplications involving humans, non-human animals (including domesticatedcompanion animals, livestock, and wildlife), and plants, includingrecreational, veterinary, agricultural, silvicultural, horticultural,and environmental applications. Other embodiments encompass diseasecontrol applications, such as controlling the spread of disease amonganimals and/or plants by controlling the vector for that disease.Exemplary vector-borne diseases of animals include, but are not limitedto: Lyme disease; Dengue Fever; Yellow Fever; tick borne-babesiosis;tuleremia; powassan-like virus infection; tick borne encephalitis;relapsing fever; malaria; encephalitis, such as the disease caused bythe West Nile Virus, Eastern equine encephalitis, St. louisencephalitis, Venezuelan equine encephalitis, Western equineencephalitis and Lacrosse encephalitis; Colorado Tick Fever;ehrlichiosis; Rocky Mountain Spotted Fever; and the Plague. Anexemplary, non-limiting vector-borne disease of plants is Dutch Elmdisease, elm yellows phytoplasmas, and apply powdery mildew arenon-limiting examples of vector-borne diseases of plants.

In any particular embodiment, the compound or composition isadministered in a pesticidally effective amount. That amount may dependon a variety of factors, including (but not limited to) the area to betreated, the pest to be treated, its metabolism, its behavior (e.g.,feeding habits, breeding, daily or seasonal activity cycles,development, nesting habits, etc.), and behavior of the host the pestinfests.

In some embodiments, the compound or composition is applied once, whilealternative embodiments employ plural applications of the same ordifferent compounds or compositions. In particular embodiments, thecompound or composition is administered on an hourly, daily, weekly,monthly, quarterly, or annual basis. In any particular embodiment, thefrequency of application may be regular or irregular, and the timeelapsed between successive applications may be the same or different.For example, and without limitation, the compounds or compositions maybe applied every eight to twelve hours; four times per day at irregularintervals; every evening; four times per week; every other day; everyother week; every other month; twice a month; every three months; everysix months; every nine months; or annually. Like the amount of thecompound or composition used in an embodiment, the frequency and numberof applications of that compound or composition may depend on a varietyof factors, including (but not limited to) the area to be treated, thepest to be treated, its metabolism, and its behavior (e.g., feedinghabits, breeding, daily or seasonal activity cycles, development,nesting habits, etc.), and behavior of the host the pest infests.

Included are embodiments where a compound or composition described aboveis applied to a particular human, non-human animal, plant, inanimateobject, or environmental locus. The compound or composition may beapplied directly to the pest, thus causing the pest to directly contactthe compound, or may be applied to some locus or host that is expectedto come into contact with the pest.

If applied to a locus, the compound may be applied to the locusgenerally, such as by an aerosol or fumigant, or applied to a human,non-human animal, plant, or inanimate object within that locus. The sizeof a particular locus may vary considerably according to the method ofapplication. For example, in area-wide applications, the compound isdispersed over a locus of an environment, rather than intentionallydirected at a particular pest, human, plant, or inanimate object. Thelocus of an area-wide application may be several hundred to thousands ofacres, if the compound is used for agricultural spraying or to controlthe spread of a vector-borne disease; in structural applications, suchas controlling pests within a home or restaurant, the locus may beseveral hundred several thousand square feet. However, in personal,veterinary, or horticultural applications, such as using topical pestrepellent spray or ointment, or using a flea shampoo to bathe a pet, thelocus may be limited to the area in the immediate vicinity of theanimal, plant, or human host.

The size of the locus also may vary according to such factors as theintended application, presence of humans or non-human animals, level ofhuman or non-human activity within the locus, type of formulationembodying the compound or composition, and environmental factors, suchas wind speed, humidity, temperature, and anticipated rainfall.

Methods of application include spraying, atomizing, dusting, immersing,coating, dressing, scattering, and pouring. In particular embodiments,the compound or composition is provided or administered to a human ornon-human animal, such as oral administration (for example, as a pill,powder, tablet, capsule, or food supplement), intravenous injection,percutaneous injection, or topical treatment. In more particularembodiments, the composition is a topical oil, lotion, or cream and thecompound is absorbed through the skin. A particular method ofapplication may be selected in accordance with the intended objectivesof and circumstances related to a particular use.

The frequency of application also may depend on the residual action ofthe particular compound or composition. “Residual action” refers to thelength of time a compound or composition may exist in a particularenvironment and remain effective. For example, one particular compoundlasts approximately 11 weeks in a protected environment before it beginsto degrade and lose effectiveness. See Example 5 below. A person using acompound, such as using a pesticidal composition having this compound asan effective ingredient to control ants, could apply the compound tosome locus in a protected environment, such as a household basement,every 11 weeks.

Some formulations embodying compounds and compositions according to anyof Formulas I-VI may offer certain advantages, such as long term effectdue to extended residual action, or high levels of safety and efficacyfor veterinary, agricultural, and nuisance pest applications.

The compounds and compositions described herein may be employed informulations intended for use in public or private homes, residences,businesses, restaurants, hospitals, or other similar places of humanactivity. In such embodiments, the formulations may be used to kill orrepel pests, such as mosquitoes, ants, spiders, or roaches, and may beapplied directly to the pests or a locus the pest is expected tocontact. For example, flea bomb or other fumigant containing an activeingredient in the form of a compound according to any of Formulas I-VIcould be used within a home, such as applied within a particular room ofa home, to control fleas. As another example, a commercial spraycontaining an active ingredient in the form of a compound according toany of Formulas I-VI could be applied to the floors and other interiorspaces of a restaurant to control cockroaches. In any such embodiment,the formulation may kill or repel a pest by directly contacting thepest, may be induced into the atmosphere of the locus, or may be appliedto a human, non-human animal, plant, or inanimate object (e.g., thesurface of a floor) expected to come into contact with the pest.

Certain embodiments employ formulations for use on humans, non-humananimals, or plants for their protection. For example, certainformulations may be insecticides and/or acaricides sprayed onto theleaves of indoor plants for controlling aphids. Other formulations mayembody compounds or compositions according to any of Formulas I-VI aslotions or oils that repel pests.

Certain embodiments encompass protection of homes, buildings, or otherstructures from nuisance insects, such as termites, cockroaches, andants. In such methods, the compound or composition may be applied to alocus within or outside the structure protected, such as spraying ontofloors or inside cupboards, or soaking the ground outside the structure.Additionally, the compound or composition may be embedded withinmaterials used to construct the structure, such as siding, wall studs,or beams.

Certain nontoxic compounds and compositions may be used to control pestsparasitic to a particular subject. The subject may be a human ornon-human animal, including domesticated animals and livestock, such asdogs, cats, birds, reptiles, cattle, swine, sheep, fowl, and goats. Insuch embodiments, the compound or composition may be provided to thehuman or non-human animal as a topical formulation, such as a cream,lotion, ointment, dip, shampoo, spotting liquid or spray, or provided inthe form of a wearable product, such as a collar, ear tag, or piece ofclothing. The compound or composition may be administered orally,rectally, or by injection, such as by a pill, solution, subcutaneousinjection, or subcutaneous implant. In any such application, thefrequency of treatment of the subject to be treated by the compound orcomposition is generally from about once per week to about once peryear, such as from about once every two weeks to about once every sixmonths, or from about once per month to about once every three months.The appropriate dose provided or administered in a particular embodimentmay vary according to the efficacy of the particular compound orcomposition; intended biocidal spectrum of the compound or composition;the physiological state or health of the subject, including allergicindications of the subject; and environmental considerations, such asexposure to wind, rain, heat, or cold. Suitable doses include from about1 to 500 mg/kg (mg of compound or composition per kg body weight of thehost), such as from about 1 to about 100 mg/kg, from about 1 to about 50mg/kg, from about 5 to about 50 mg/kg, from about 5 to about 10 mg/kg,from about 10 to about 100 mg/kg, or less than 1 mg/kg.

The compound or composition may be used to clean the animal, such as byan owner bathing or placing a flea collar on a pet, or in veterinaryapplications. Cleaning an animal may be distinguished from treating ananimal body, since an animal in good health would not requiresubstantial treatment to correct a deficiency of health.

In certain embodiments, the compounds or compositions applied in anarea-wide manner, such as in protection of agricultural crops describedbelow. In addition to agricultural applications, area-wide applicationsmay include silvicultural, horticultural, or other forms ofenvironmental pest management and control. In such embodiments, acompound or composition may be applied to plant foliage, such asspraying or dusting, or applied to the soil, such as drenching aparticular locus with a liquid formulation or applying the activeingredient in solid form to a locus. In some instances, plants within oradjacent to the locus of application may absorb the active ingredient orcomposition through their roots. In other instances, the activeingredient will remain in the environment, such as when a compound orcomposition is applied to a stagnant body of water to control mosquitolarvae.

Certain embodiments use the compounds and compositions described hereinfor pest control in food production and storage. For example, certaincompositions may be used as agricultural pesticides to control pests andprotect grain, vegetable, herb, spice, or fruit crops. Compositions alsomay be used to control pests affecting other plants useful or importantin agricultural or horticultural production, such as those plants orcrops producing cotton, flax, tobacco, hemp, rubber, nuts, nurserystock, and ornamental plant parts.

The compounds and compositions according to any of Formulas I-VI may beused to protect plant products not only during growth and production,but also during storage or transport of such products. For example, someembodiments use compounds or compositions to protect grain stored insilos, bales of cotton or tobacco stored in warehouses, or bushels offruit being transported from an orchard.

The compounds and compositions described herein also may be used toprotect plant propagation material, such as seeds, fruit, tubers, orplant cuttings. The propagation material may be treated with theformulation before planting, such as soaking, coating, or dressing seedsprior to sowing. The compounds and compositions also may be applied tothe soil where the propagation material will be planted, such asin-furrow application to protect seeds.

In such applications, the compound or composition may be applied toprovide a certain concentration of the compound in the environment at aparticular locus. That certain concentration may be measured,established, or determined according to the needs of the user. Forexample, when applying the compound or composition to crops, the rate ofapplication may depend on the nature of soil, the type of application(e.g., spraying crop foliage, burial in soil), the crop plant to beprotected, the pest to be controlled, the prevailing climaticconditions, growing season, proximity to residential areas or protectedenvironments, and other factors. As another example, when applying thecompound or composition to stored or transported agricultural products,the rate of application may depend on the localized environment (e.g.,storage within a warehouse, storage under a covered shelter, transportwithin a trailer), expected duration of storage, product to beprotected, the pest to be controlled, economic considerations, and otherfactors. In certain embodiments, the rates of concentration are in therange from about 0.01 to about 1000 ppm (parts-per-million), such asfrom about 0.1 to about 500 ppm, of active ingredient. In area-wideapplications, rates of application per hectare may be from about 0.5g/ha to 2000 g/ha, such as particularly from about 10 to 1000 g/ha, orfrom about 20 to 600 g/ha. As one non-limiting example, pesticides forthe control of mosquito vectors of malaria may be used in area-wideapplications at a rate of application of about 70 g/ha to about 1.15kg/ha.

Use of pesticides is regulated in the United States by state and federalagencies, including the Environmental Protection Agency (EPA) and Foodand Drug Administration (FDA). Relevant regulatory programs include theFederal Insecticide, Fungicide and Rodenticide Act (FIFRA) and theFederal Food, Drug and Cosmetic Act (FD&C Act). Certain articles ofmanufacture in accordance with these governmental and regulatoryconsiderations may be made using compounds or compositions according toany of Formulas I-VI.

In such embodiments, a pesticidally active compound according to any ofFormulas I-VI is embodied in an acceptable carrier and stored within acontainer capable of storing the composition for its shelf life. Thecontainer may be made of any suitable material such as plastic or otherpolymer, glass, metal, or the like. Printed instructions and/or aprinted label indicating that the composition may be used to controlpests are associated with this container. The instructions and/or labelmay provide information regarding the use of the composition forpesticidal purposes in accordance with the treatment method set forthherein and may be associated with the container by being adhered to thecontainer, or accompanying the container in a package. The label mayindicate the composition is approved for use as a pesticide, and theinstructions may specify the pests intended to be controlled by thecomposition, the method and rate of application, dilution protocols, useprecautions, and the like. Additionally, the container may include afeature or device for applying the composition to the pest population orlocus to be treated. For example, if the article of manufacture includesa liquid composition, the feature or device may be a hand-operated,motorized, or pressurized pressure-driven sprayer. In certainembodiments, the article of manufacture includes, packaged together, avessel—such as a tube, barrel, bottle, bottle, or can—containing thecomposition and instructions for use of the composition for controllinga pest. In other embodiments, the article of manufacture is a devicethat includes the compound as part of the device, such as a surfacecoated with the compound, for example a bait trap or flea collar. Inalternative embodiments, the article of manufacture includes packagingmaterial containing the composition. Additionally, the packagingmaterial may include a label indicating that the composition may be usedfor controlling a pest and, in particular embodiments, a pesticide forkilling a pest. Examples of articles of manufacture include, but are notlimited to, spray bottles of a ready-to-use formulation for householduse; bottles, cans, or barrels containing concentrated formulations thatmay be diluted for area-wide applications; containers of concentratedformulations for use in industrial settings; flea collars or ear tagsfor domesticated companion animals and livestock; bottles or kits forshampooing, dipping, or cleaning domesticated companion animals orlivestock; a bottle containing a formulation for human use as a shampooor body wash; plastic tubules containing a topical oil for applying to adomesticated animal; and rodent bait boxes or host targeted bait boxescontaining a pesticidal composition for killing ectoparisites infestingthe host animal.

EXAMPLES

The following examples are provided to illustrate particular features ofcertain embodiments. The scope of the invention should not be limited tothose features exemplified.

Example 1 Isolating Certain Compounds from Alaska Yellow Cedar

This example illustrates one method of isolating pesticidal ermorphilanesesquiterpenes from Alaska yellow cedar (Chamaecyparis nootkatensis),including nootkatone, 13-hydroxy-valencene, and valencene-11,12-epoxide.

Alaska Cedar Tree

An Alaska cedar tree was collected from the Hunqry Mountain area on theSol Duc River drainage on the western slopes of the Olympic Mountains inthe Olympic National Forest. A botanical voucher specimen (#188046) isdeposited in the Oregon State University Herbarium. The heartwood wasseparated from sapwood and bark and then chipped in a grinder toapproximately 15×10 mm chips and stored at room temperature until used.

Extraction of the Essential Oils

Steam Distillation

Steam distillation of the heartwood chips was carried out in a standardapparatus, in which 1.5 kg of chips were steam distilled for 6-12 hoursto yield 26 grams of essential oil. The oil was recovered by extractionof the combined water/oil distillate with diethyl ether. The diethylether solution was dried over anhydrous sodium sulfate and evaporated ona rotary evaporator under reduced pressure, resulting in a yellow oil.Nootkatin tended to crystallize out in the condenser duringdistillation, so diethyl ether was periodically used to dissolve thesecrystals into the oil fraction. Nootkatin crystallized out of the Alaskacedar oil when it was placed in the refrigerator. These crystals wererecovered by decanting the oil and re-crystallization of the nootkatinfrom the oil solution to give pure material. The remaining Alaska cedaroil that was used in this study was substantially free of nootkatin withonly trace amounts of nootkatin present.

Extraction by Diethyl Ether

200 grams of Alaska cedar heartwood chips were twice extracted with 3liters of diethyl ether for 24 hours at room temperature to ensurecomplete extraction of the oil. The combined ether solution wasfiltered, dried with anhydrous sodium sulfate, and evaporated to give2.2 grams of an oil.

Isolation of Compounds

Isolation from the Steam Distilled Oil:

The essential oil components were separated and purified by traditionalcolumn chromatography. When packing a column, a degased slurry ofsolvent and adsorbent (Kieselgel 60 PF₂₅₄ Silica gel, Germany) waspoured into a glass column with a diameter and height determined bysample size. The solvent was drained until its level was just over thetop of adsorbent. The stopper of the column was closed and the columnwas ready for use.

The distilled oil (62 grams) was dissolved in 50 ml of hexane andchromatographed over a Silica gel 60 column (7×45 cm) using a gradientsolvent mixture of hexane and diethyl ether from 100% hexane to 60:40(hexane/diethyl ether, v/v). Aliquots of 20 mL eluent were collectedwith a Gilson FC-100 fraction collector and monitored by TLC developedwith dichloromethane. The plates were visualized under UV light andsubsequently sprayed with acidic vanillin solution, followed by heating.Aliquots of eluent with same component checked by TLC were combinedtogether to form one fraction. Seven major fractions were obtained afterthe first chromatographing of the crude distilled oil (62 g): I (11.31g), II (20.6 g), III (0.48 g), IV (18.03 g), V (3.45 g), VI (5.08 g),VII (1.13 g).

Fraction I was found to mainly contain valencene and nootkatene and atrace of methyl carvacrol by gas chromotography.

Fraction II was highly pure carvacrol checked by gas chromotography.

Fraction III was a mixture of trace components by comparing itschromatogram to that of the crude oil in gas chromotagraphy.

Fraction IV showed two main spots on a TLC plate developed bydichloromethane, one visible under UV light and the other only afterbeing sprayed with acidic vanillin solution, followed by heating. TheirRf values were 0.29 and 0.42, respectively. This fraction was analyzedby gas chromotography and found to consist of nootkatone (Rf 0.29) andone unknown compound named “unknown compound 1,” Rf 0.42. A portion ofthis fraction (5 g) was rechromatographed twice over a Kieselgel column(5×45 cm) with dichloromethane as mobile phase and yielded the two purecompounds, nootkatone (1.55 g) and unknown compound 1 (0.68 g). As shownin FIG. 1A, unknown compound 1 was subsequently identified as13-hydroxy-valencene.

Fraction V was still a mixture, which contained small amount of almostevery component in the crude oil.

Fraction VI checked by gas chromotography and TLC was found to containone main compound (Rf 0.43 in hexane/ethyl acetate 70/30 v/v). Thisunknown compound 2 (8 mg) was yielded from one portion of this fraction(30 mg) after preparative HPLC procedures and, as shown in FIG. 1A,later identified as nootkatol.

Fraction VII contained highly pure unknown compound 2 (nootkatol)checked by gas chromotography.

A graphic representation of separation of these fractions is illustratedin FIGS. 1A and 1B.

Isolation from the Diethyl Ether Extract:

The diethyl ether extract (2.5 g) was chromatographed on a Silica gelcolumn (5×44 cm) with diethyl ether and dichloromethane as the gradientsolvent system from 90:10 (dichloromethane:diethyl ether, v/v) to 100%diethyl ether. See FIG. 1. Another unknown compound, named “unknowncompound 3,” was obtained as Fraction 2. As shown in FIG. 1B, unknowncompound 3 was later identified as valencene-11,12-epoxide.

Spectroscopic Data for Isolated Compounds

Eremophil-1(10),11-dien-13-ol. Isolated from Fraction IV of the crudeessential oil extraction and originally listed as “unknown compound 1”but later identified as eremophil-1(10),11-dien-13-ol. See FIG. 1A. Paleyellowish oil. MW=220. High Resolution MS revealed mass (220.18271) andformula (C₁₅H₂₄O). C₁₅H₂₄O requires 220.18272. [α]₅₈₉+61.9 (C 2.26 inchloroform). Rf 0.42 (in dichloromethane). MS (70 ev), m/z 220([M⁺]100), 202(40), 189(81), 161(77), 105(67), 91(65), 79(54). ¹³C NMR (ppm):154.5, 143.2, 120.8, 108.3, 65.7, 45.8, 41.3, 38.3, 37.1, 34.0, 33.1,27.5, 26.3, 18.8, 16.0. ¹H NMR (δ): 0.87 (3H, d, J=6 Hz), 0.95 (3H, s),1.01 (1H, d, J=12.6 Hz), 1.21 (1H, dd, J=4.1, 13.3 Hz), 1.41 (3H, m),1.59 (1H, m), 1.82 (1H, dm?), 1.93 (1H, m), 2.01 (2H, m), 2.09 (1H, ddd,J=14.0, 4.16, 2.66 Hz), 2.31 (1H, tt, J=12.7, 3.0 Hz), 4.12 (2H, s),4.88 (1H, s), 5.02 (1H, s), 5.33 (1H, t, J=2.4 Hz), 7.26, (1H, s).

Nootkatone. Pale yellowish oil. MW=218, C₁₅H₂₂O. [α]₅₈₉+152 (1.51 inchloroform). Rf 0.29 (in dichloromethane). ¹³C NMR (ppm): 199.7, 170.6,149.0, 124.6, 109.2, 43.9, 42.0, 40.4, 40.3, 39.3, 33.0, 31.6, 20.8,16.8, 14.8. ¹H NMR (δ): 5.77 (1H, s), 4.74 (2H), d), 1.74 (3H, s), 1.13(3H, s), 0.97 (3H, s).

Nootkatol. Colorless oil. MW=220. High Resolution MS revealed mass(220.18165) and formula (C₁₅H₂₄O). C₁₅H₂₄O requires 220.18272.[α]₅₈₉+41.3 (C 1.52 in chloroform). R_(f)0.43 (in hexane/ethyl acetate),0.4 (in hexane/diethyl ether 50:50 v/v), 0.74 (indichloromethane/diethyl ether 50:50 v/v). MS (70 ev), m/z 220([M⁺] 100),203(21), 187(4), 177(60), 162(6), 138(13), 121(20), 107(21), 93(22),81(16), 67(13). ¹³C NMR (ppm): 150.6, 146.5, 124.7, 108.9, 68.4, 45.0,41.2, 39.7, 38.6, 37.6, 33.3, 32.8, 21.2, 18.6, 15.8. ¹H NMR (δ): 0.89(3H, d, J=6.9 Hz), 0.95 (1H, J=2.7 Hz?), 0.99 (3H, s), 1.20 (1H, dm,J=4.3 Hz), 1.37 (1H, td, J=12.4, 10.0 Hz), 1.51 (1H, m, J=2.1), 1.71(3H, s), 1.76 (1H, td, J=2.0, 6.5 Hz), 1.79 (1H, dd, J=2.0, 4.5), 1.85(1H, dd, J=12.6, 2.7 Hz), 2.1 (1H, ddd, J=14.1, 4.2, 2.6 Hz), 2.25 (1H,tt, J=12.4, 3.0 Hz), 2.33 (1H, m), 4.25 (1H, m), 4.68 (2H, m), 5.32 (1H,d, J=1.6 Hz).

valencene-11,12-epoxide. Isolated from Fraction 2 of the diethyl etherextract and originally listed as “unknown compound 3,” but lateridentified as valencene-11,12-epoxide. See FIG. 1B. Dark yellow oil.MW=220. High Resolution MS revealed mass (220.18280) and formula(C₁₅H₂₄O). C₁₅H₂₄O requires 220.18272. [α]₅₈₉+58.5 (C 1.17 inchloroform). R_(f)0.36 (in dichloromethane/diethyl ether 50:50 v/v). MS(70 ev), m/z 220([M⁺] 85), 189(74), 178(6), 161(100), 135(41), 121(25),107(38), 81(42), 75(44). ¹³C NMR (ppm): 143.4, 120.5, 75.0, 69.0, 41.5,40.4, 39.9, 38.1, 32.9, 29.6, 27.6, 26.3, 20.4, 18.8, 16.1. ¹H NMR (δ):0.86 (1H, m), 0.89 (3H, d, J=6.27 Hz), 0.93 (3H, s), 1.00 (1H, dd,J=4.7, 13.2 Hz), 1.07 (3H, s), 1.42 (3H, m), 1.71 (1H, ddd, J=2.6, 4.7,12.2 Hz), 1.84 (1H, tt, J=3.0, 12.6 Hz), 1.98 (3H, m), 2.07 (1H, ddd,J=2.6, 4.2, 14.1 Hz), 2.27 (1H, m), 3.43 (1H, d, J=10.14 Hz), 3.59 (1H,d, J=11.28 Hz), 5.32 (1H, t, J=2.5 Hz).

NMR experiments were run on a Bruker Model AM 400 spectrometer with theXWINNMR software package, using CDCl₃ as the solvent and TMS as aninternal standard for chemical shifts given in ppm. DEPT (DistortionlessEnhancement by Polarization Transfer) experiments were performed usingboth pulses of 135° C. and 90° C. ¹H-¹H COSY, ¹H-¹³C HSQC, HMBC, NOEswere also performed on the instrument according to the standardprocedures described by Bruker. EI-MS was done with a Kratos MS-50TCmass spectrometer.

A gas chromatograph (GC-17A Shimadzu, Japan) was used for monitoringcomposition of fractions and identifying pure compounds by usingstandards. The gas chromatograph was equipped with a flame ionizationdetector (FID). The column (30 m×0.25 mm DB-5, 0.25 μm, J&W Scientific)was temperature programmed from 100° C. for 1 minute, then to 150° C. ata rate of 5° C./min, then to 220° C. at 3° C./min, and finally to 240°C. at 5° C./min and held at that temperature for 2 minutes.

GC-MS analysis was carried out on a HP 5972 GC/MS to confirm thosepreviously known compounds in Alaska cedar heartwood oil. One microliterof a 587 ng/μl solution of the distillate dissolved in hexane wasinjected into the injector maintained at 250° C. A 30 m×0.25 mm ID DB-5column was used and temperature programmed from 50° C. initially heldfor 5 minutes to 300° C. finally at a rate of 5° C./min. The transferline temperature was 280° C. The MS was operated in electron impact modewith a 70 eV ionization potential and was scanned from 50-560 m/z.

The optical rotations were measured on a digital polarimeter (JASCO,MODEL DIP-370, Japan) with a Na lamp (589 nm) as the light source.Chloroform was used as the solvent.

Analytical thin-layer chromatography (TLC) was performed on aluminumplates pre-coated with Kieselgel 60 F₂₅₄ (EM, Germany) to monitor thecourse of column separation and act for a preliminary guideline toselect mobile phase for column separation.

The solvent systems used for TLC analysis were:

-   -   (1) Hexane:Ethyl acetate (70:30 v/v)    -   (2) Dichloromethane    -   (3) Hexane:Diethyl ether (50:50 v/v)        The spots on TLC plates were visualized under UV light and        sprayed with acidic vanillin solution (1 g vanillin, 50 mL        absolute EtOH, and 10 mL concentrated HCl), followed by heating.

Preparative HPLC was performed on a Waters Millipore Model 510 systemusing a normal phase column (Silica-prep, 250×10 mm 10 μm, Phenomenex).Degased hexane (A) and ethyl acetate (B) were used as the gradientmobile phase, starting from 15 up to 30% B in 20 min, to 40% B in 30min. The total flow rate was maintained at 3.5 mL/min. An UV detector(Lambda-Max Model 481 LC spectrophotometer, Waters Millipore) wasconnected to the outlet of the column and operated at 254 nm and 0.01AUFS. A computer-based data system (Maxima 820) was connected to thesystem for monitor and control. Fractions were collected according tothe peaks shown on the screen.

A Buchi Rotavapor Model R-110 equipped with a Buchi 461 Water Bath wasused for the removal of solvent from the samples under reduced pressureby using a water aspirator. The temperature of water in the bath wasmaintained at 30° C.

All solvents used were ACS grades and re-distilled prior to use. Allwater was also distilled before use.

Example 2 Pesticidal Properties of Certain Compounds on Ticks and Fleas

Samples of the compounds listed below were obtained from Dr. Karchesy, aco-inventor, and screened for biocidal activities against nymphal Ixodesscapularis. In this example, 2% acetone solutions (wt./vol.) of thecompounds were applied to inner surfaces of 2-dram friction cap vials.Vials and caps were treated then allowed to dry for a minimum of 4 hoursbefore placing 10 nymphs in each container. These same vials were usedto challenge additional nymphs and adult I. scapularis ticks and adultXenopsylla cheopis fleas through five weeks to observe any possibleresidual activity. Results of this biocidal screening are presented inTable 5.

TABLE 5 I. scapularis I. scapularis nymphs nymphs I. scapularis X.cheopis Compound (24 h) (72 h) adults (4 wk) adults (5 wk) Valencene10/10 0/5 0/5 0/5 Nootkatene 10/10 4/5 0/5 0/5 Nootkatone 10/10 5/5 5/55/5 valencene- 10/10 5/5 5/5 5/5 11,12-epoxide Nootkatin  1/10 0/5 0/50/5

Numbers in parentheses refer to length of treatment in terms of hours(h) or weeks (wk). Data is presented in terms of number killed/numbertested. For test periods longer than 24 hours, the treated vials wereallowed to sit for the stated period after drying (72 h, 4 wk, or 5 wk)and each group of arthropods was added to the vials for the final 24 hof the test period.

These bioassays demonstrate that four of the five compounds had biocidalactivity against ticks and fleas and that nootkatone andvalencene-11,12-epoxide were the most efficacious and persistent.

Example 3 Pesticidal Properties of Certain Compounds on Mosquito

Using a bioassay method similar to that presented in Example #2, thesusceptibility of Aedes aegypti adults to the five compounds presentedin Example #2 at 24 hours challenge was determined to be 100% mortalityexcept for nootkatin (20% mortality).

Example 4 Pesticidal Properties of Nootkatol on Certain Arthropods

Nootkatol was tested using the bioassay presented in Example 2. Thebiocidal activity of nootkatol was determined to be essentiallyequivalent to the activity of nootkatene against ticks, fleas andmosquitoes.

Example 5 Persistence of Biocidal Activity of the Compounds

Persistence of biocidal activity was determined for nootkatone andvalencene-11,12-epoxide by using the vials as treated with the 2%solutions of Example 2. This bioassay employed groups of ticks, fleas,formicids and termites using a method similar to that presented inExample 2. Each group of arthropods demonstrated 100% mortality after 24hour exposure at each test through 10 weeks for both chemicals. At 11weeks, the biocidal activity began to dissipate below the 100% mortalitylevel.

Example 6 Comparison of Extracted Nootkatone Samples

Two nootkatone samples were compared to nootkatone isolated from Alaskayellow cedar using the method presented in Example 1. The first samplewas a natural extract of nootkatone taken from grapefruit oil purchasedcommercially. The second sample was a synthetically produced nootkatonepurchased commercially.

Both samples were compared to the nootkatone originating from Alaskayellow cedar in terms of their biocidal activities. Using a bioassaymethod similar to that presented in Example 2, no differences werediscernible among the nootkatone samples in terms of their biocidaleffect on ticks, fleas and mosquitoes.

Example 7 Effectiveness of Compounds Against Ticks

Baseline dose-mortalities were established for nymphal I. scapularis fornootkatone and valencene-11,12-epoxide using the method described inMaupin, G. O., and Piesman, J., J. Med. Entomol., 31: 319-21 (1994).

A comparison of relative potency was made with published data forcarbaryl and permethrin (Maupin and Piesman, 1994) and Alaska yellowcedar essential oil (Panella et al., J. Med. Entomol. 34: 340-45 (1997)is presented in Table 6.

TABLE 6 Compound LD₅₀ Essential oil of Alaska 151.0 × 10⁻³  yellow cedarNootkatone 4.2 × 10⁻³ 13-hydroxy-valencene 4.1 × 10⁻³ Carbaryl 7.2 ×10⁻³ Permethrin 3.0 × 10⁻³

Nootkatone and valencene-11,12-epoxide were extracted from Alaska yellowcedar essential oil. Both compounds were approximately 50 times morepotent and about 98% more effective against nymphal ticks than theirparent source, essential oil of Alaska yellow cedar. While permethrinand carbaryl demonstrated greater effectiveness than these two biocidalsesquiterpenes, this relative potency is not absolutely accurate sincethe permethrin and carbaryl samples were technical grade (>99% pure),while the two biocidal sesquiterpenes were extracted at a 90-95% puritylevel.

Example 8 Effectiveness of Compounds Against Mosquito

The susceptibility of mosquitoes to valencene-11,12-epoxide wasdetermined using a method similar to that presented in Example 2. Culexp. pipiens was treated with serial dilutions of valencene-11,12-epoxidefrom 0.125% down to 0.0045%. Due to the extreme sensitivity of thisspecies to the biocidal activity of valencene-11,12-epoxide, thecorresponding LD₅₀ could not be calculated by Probit analysis. At thelowest tested dosage, the mortality rate was still 64%. Therefore, theLD₅₀ for compound 10 against C. pipiens is <4.5×10⁻³. Similar resultswere obtained when both valencene-11,12-epoxide and nootkatone wereassayed for their biocidal activities against Aedes aegypti, thusdemonstrating that mosquitoes are quite sensitive to these compounds.Table 7 compares the effectiveness of raw essential oil of Alaska yellowcedar, nootkatone, and valencene-11,12-epoxide against two mosquitoes,A. aegypti and C. pipiens.

TABLE 7 Compound A. aegypti LD₅₀ C. pipiens. LD₅₀ Essential oil ofAlaska   32.0 × 10⁻³   61.0 × 10⁻³ yellow cedar Nootkatone  <4.5 × 10⁻³ <4.5 × 10⁻³ valencene-11,12-epoxide  <4.5 × 10⁻³  <4.5 × 10⁻³

Example 9 Effectiveness of Compounds Against Ticks, Fleas, andMosquitoes

The pesticidal activities of valencene-11,12-epoxide,valencene-13-aldehyde, nootkatone-1,10-epoxide, andnootkatone-1,10-11,12-diepoxide against ticks, fleas, and mosquitoeswere assayed.

Materials and Methods.

Plant Extracts. All compounds were produced in the Forest ChemistryLaboratory at Oregon State University (Corvallis, Oreg.) from an Alaskayellow cedar (Chamaecyparis nootkatensis) specimen collected under aspecial collection permit from the United States Forest Service. A plantvoucher specimen (#188046) was deposited at Oregon State UniversityHerbarium (Corvallis, Oreg.).

Some tested compounds—carvacrol, nootkatin, nootkatene, valencene,nootkatone, nootkatol, and 13-hydroxy-valencene—were isolated from thesteam distilled essential oil of Alaska yellow cedar heartwood (seeExample 1). Valencene-11,12-epoxide was isolated from the diethyl etherextract of the ground heartwood. See Xiong, Y., Essential oil componentsof Alaska cedar heartwood (Masters Thesis, Oregon State University,Corvallis, Oreg., 2001). Valencene-13-aldehyde was prepared by oxidationof valencene with SeO₂. Nootkatone-1,10-epoxide andnootkatone-1,10-11,12-diepoxide were prepared from nootkatone with H₂O₂,and nootkatone-11,12-epoxide was prepared using m-chloroperbenzoic acid.Commercial samples of nootkatone for comparison were obtained fromBedoukian Research, Inc., Danbury, Conn. (semi-synthetic crystalline)and Frutarom, Inc. N.J. (from grapefruit oil). As shown in Table 8, eachcompound was assigned a numerical identifier for this Example (theidentifiers for the compounds used in this Example may be different thanthe identifiers used elsewhere in this application as specified in Table1 above). The compounds identified by an asterisk—nos. 1, 2 and3—demonstrated no pesticidal activity after 24 hours exposure.

TABLE 8 Compound names, numerical identifiers, and source for compounds.No. Compound name Source 1 3-Carene* Alaska yellow cedar, 2Terpinen-4-ol* Alaska yellow cedar, 3 Methyl carvacrol* Alaska yellowcedar, 4 Carvacrol Alaska yellow cedar, 5 Valencene Alaska yellow cedar,6 Nootkatene Alaska yellow cedar, 7 Nootkatone, crystalline Alaskayellow cedar, 8 Nootkatone Grapefruit oil 9 Nootkatone Synthetic 1013-hydroxy-valencene Alaska yellow cedar, 11 Nootkatin Alaska yellowcedar, 12 Nootkatol Alaska yellow cedar, 13 Valencene-11,12-epoxideAlaska yellow cedar, 15 Valencene-13-aldehyde Valencene 16Nootkatone-11,12-epoxide Nootkatone 17 Nootkatone-1,10-epoxideNootkatone 18 Nootkatone-1,10-11,12- Nootkatone diepoxide

Tick colonies. Nymphal I. scapularis ticks (8-12 weeks old) were used inall trials and were obtained from the F₁ offspring of adult I.scapularis ticks collected in Bridgeport, Conn. There was no knownpesticide used at this location. Ticks were maintained at 21° C., 90%RH, and received a 16:8 h (light:dark) cycle as described in Piesman,J., J. Med. Entomol. 30:199-203 (1993).

Flea Colonies. Adult X. cheopis fleas (1-3 wk old) were obtained from acolony founded more than eight years ago using adults received from TomSchwan, Rocky Mountain Laboratories, Hamilton, Mont. The area from whichthese original adult fleas were obtained, and the resulting colony, haveno known histories of pesticide exposure. Colonies were maintained inglass jars containing a 4:1:1:1 ratio of sawdust, dried beef blood,powedered milk, and powdered mouse chow and were held at 23° C., 85% RH,and received a 24 h dark cycle.

Mosquito Colonies. Aedes aegypti adult mosquitoes were obtained from anexisting colony at the Centers for Disease Control and Prevention (CDC),Division of Vector-Borne Infectious Diseases (DVBID), Fort Collins,Colo. This colony has been maintained for over fifteen years with noknown history of exposure to pesticides. Mosquitoes were reared at 28°C., 85% RH, and a 14:10 h (light:dark) cycle. Larvae were grown inde-ionized water and fed ground liver powder solution ad libitum. Fourthinstars were removed and placed in emergence cages, and adults were feda 2% sucrose solution until assayed. Adults were exposed to testproducts at 4-7 d after emergence.

Tick and Flea Bioassays. Concentrations of the compounds were preparedby 2-fold serial dilutions of a 0.5% (wt:vol) solution of the extractsin acetone. The approximate toxicity of individual compounds wasdetermined with a total of 8 doses ranging from 0.002% to 0.25%. All 13compounds were run in duplicate with more active compounds replicatedonce. A control treated with acetone only was run with each series. Tickand flea susceptibility was evaluated using a modified disposablepipette method. See, Barnard, D. R., et al., J. Econ. Entomol. 74:466-69(1981).

Groups of 10 nymphal I. scapularis and X. cheopis fleas were used in alltests, resulting in a total of 5,240 nymphs and 4,860 fleas exposed tothe compounds. The inner surface of 2-dram friction cap vials weretreated with the compound/acetone solution, or acetone only as acontrol, and left to air-dry overnight. Three to 4 holes were made inthe plastic cap to allow for air exchange. Nymphal ticks and adult fleaswere then placed directly into the vials using forceps. Vials containingticks or fleas were placed in desiccators for 24 h at 21° C. and 90% RH.Morbidity and mortality was recorded at 1, 2, 4, 8, and 24 h afterinitial exposure.

After 24 h, ticks were considered alive if they exhibited normalbehavior when breathed upon or physically stimulated with wooden dowels.For each time point, if ticks were incapable of movement, maintainingnormal posture, leg coordination, ability to right themselves, or anysigns of life, they were considered moribund or dead. Results from testswhere more than 10% of the control population died were discarded andretested.

Efficacies of individual compounds were determined by calculating lethalconcentration 50% (LC₅₀) and 90% (LC₉₀) wt:vol by probit analysis usingthe LdP Line software (copyright 2000 by Ehab Mostafa Bakr), availablevia the Internet.

Mosquito Bioassay. Adult mosquitoes were tested using the bottlebioassay method of Brogden, W. G., and McAllister, J. C., J. Am. Mosq.Control Assoc. 14: 159-64 (1998), with minor modifications. Naturalproduct extracts were two-fold serially diluted for a total of 8concentrations ranging from 0.002% to 0.25% in 1.5 ml of acetone.Individual dilutions were added to 250 ml Wheaton glass bottles andcapped. Bottles were manipulated to evenly coat all inner surfaces. Thecaps were removed and bottles were allowed to air-dry overnight. Oncebottles were completely dry, 10-50 adult mosquitoes were aspirated intothe bottles. Mosquitoes were held in the bottles for 24 h at 23° C. and85% RH. Morbidity and mortality were recorded at 15, 30, 45, and 60 minand 24 hour intervals. One bottle for each replicate was treated withacetone only and served as a control. The dose-mortality data wasevaluated with probit analysis, using the LdP Line software, todetermine LC₅₀ and LC₉₀ values.

Residual Activity. To determine persistence of pesticidal activity ofthe compounds, 2-dram friction cap vials and 250 ml Wheaton glassbottles were treated using the same series of dilutions as describedabove. Treated vials and bottles, minus vector test species, were heldat 21° C. with the ability of air exchange to take place (3-4 holes inthe lids of 2-dram vials and lids on Wheaton bottles loosely applied).On day 7 after treatment, 2-dram vials were loaded with 10 ticks or 10fleas and Wheaton bottles with 10-50 mosquitoes and held as previouslydescribed. Morbidity and mortality data for each test subject wasrecorded using the same time points. Any extracts that displayedacaricidal/insecticidal activity were retested at 2 and 4 wk aftertreatment. Dose mortality data was evaluated using probit analysis viathe LdP Line software.

Results.

The susceptibility of I. scapularis nymphs, X. cheopis adults, and Ae.aegypti adults are presented in Tables 9-11 below. The terpenes,compounds 1-3, were found to be ineffective against all three arthropodsin initial screenings, and, therefore were not further analyzed. Thefourth terpene, carvacrol (compound 4), exhibited significant biologicalactivity against ticks, fleas and mosquitoes with LC₅₀ values of 0.0068,0.0059, and 0.0051 respectively.

Compounds 5-12 are eremophilane sesquiterpenes. Compound 5, valencenewas effective only against mosquitoes and demonstrated an LC₅₀ of 0.015.Nootkatene, compound 6, was effective against all three pest species,demonstrating LC₅₀ values of 0.011 for ticks, 0.017 for fleas, and 0.027for mosquitoes after 24 h. This compound did not demonstrate anyresidual activity after one week. The 3 preparations of nootkatone andβ-hydroxy-valencene demonstrated the greatest activity in terms of LC₅₀and LC₅₀ of the eremophilane sesquiterpenes. Compound 7, nootkatone fromAlaska yellow cedar, was most effective against nymphal I. scapularis,with an observed LC₅₀ value of 0.0029. Adult X. cheopis were mostsusceptible to the natural grapefruit extract of nootkatone (compound 8)with an LC₅₀ value of 0.0029. Finally, compound 10(13-hydroxy-valencene) proved to be the most toxic to adult mosquitoes(LC₅₀ of 0.0034).

Although there were slight differences in susceptibility to compounds7-10, depending on the vector species, overall LC₅₀ and LC₉₀ values didnot differ significantly. Of the last two compounds tested in theeremophilane sesquiterpene ring system (compounds 11 and 12), onlycompound 12 (nootkatol) exhibited biological activity against all threespecies. The LC₅₀ values were slightly greater for ticks (LC₅₀=0.023)and fleas (LC₅₀=0.024), but comparable to nootkatone against mosquitoes(LC₅₀=0.004).

The remaining five compounds (compounds 13 and 15-18) are derivatives ofnootkatone and valencene. See Table 8. Compound 13, an epoxide, showedsome activity against mosquitoes only (LC₅₀=0.295). Compounds 17 and 18(nootkatone-1,10 epoxide and nootkatone diepoxide) had the greatestactivity against fleas (LC₅₀=0.017 for compound 17 and LC₅₀=0.064 forcompound 18) and ticks (LC₅₀=0.02 for compound 17 and LC₅₀=0.061 forcompound 18). Valencene-13-aldehyde (compound 15) demonstrated LC₅₀ andLC₉₀ values equivalent to those of nootkatone and 13-hydroxy-valencene(compound 10): LC₅₀=0.0059 against ticks, LC₅₀=0.0049 against fleas andLC₅₀=0.0024 against mosquitoes.

Compounds were analyzed for residual activity based on results obtainedfor their 24 hour activities. A total of six compounds were examinedwith five demonstrating residual activity for at least four weeks, asshown in Tables 12-14. Compound 6 was nearly inactive beyond week aftertreatment and, therefore, not tested any further.

Compounds 7, 8, 9, 10 and 15 all demonstrated considerable biologicalactivity through the four week time period. While the residual activityof compound 7 against I. scapularis nymphs remained virtually unchangedthroughout the study, compounds 8 and 9 produced the lowest LC₅₀ valuesoverall (Table 12). Compound 10 was still effective against ticks atfour weeks, though a decrease in activity compared to earlierobservations was noticed.

All five compounds remained active against fleas through the four-weekperiod, with compound 7 producing the lowest LC₅₀ values (Table 13).Measurable residual activity was also observed in mosquitoes for all 5compounds. Interestingly, the effectiveness of compounds 7 and 10increased over the four week period (demonstrated by decreasing LC₅₀values), while activity decreased for compounds 8 and 9 (Table 14).Compound 7 had the lowest LC₅₀ values after 4 wk against ticks(LC₅₀=0.026) and fleas (LC₅₀=0.031).

These dose-mortality results indicate that these compounds—such asnootkatone, carvacrol, 13-hydroxy-valencene, andvalencene-13-aldehyde—function as effective pest control agents andpesticides. These compounds have the ability to knock down insectsquickly and maintain a comparable level of activity for several weeks.

These individual compounds isolated from Alaska yellow cedar oil aremore effective pest control agents than the crude oil itself, based onthe LC₅₀ values observed for the crude oil against ticks (LC₅₀=0.151),mosquitoes (LC₅₀=0.032), and fleas (LC₅₀=0.337). These values areseveral times greater than what was observed for nootkatone, carvacrol,13-hydroxy-valencene and valencene-13-aldehyde.

Furthermore, residual activity of the crude oil decreased rapidly afterthe initial treatment and was undetectable after 21 days against nymphalI. scapularis, indicating that the individual compounds obtained fromthe crude oil, and derivative compounds, are more stable and lessvolatile over time than the crude oil.

The methods used in these Examples are representative of types ofapplications occurring outside a laboratory setting. In most suchapplications, the pest would directly contact the compounds as they doin the bottles and vials, rather than ingesting the compound as a feedsupplement, thus replicating conditions existing in a field setting. Forexample, the compound or composition might be applied to the walls ofdwellings or stagnant pools of water to control mosquitoes, or it wouldbe applied to vegetation or domesticated animals to control ticks andfleas.

TABLE 9 Response of I. scapularis nymphs after 24 h exposure. (95% CI inparentheses; LC50 and LC90 values expressed in terms of percentconcentration, wt:vol) Com- pound LC₅₀ LC₉₀ Slope 4 0.0068(0.0054-0.0084) 0.014 (0.011-0.022) 3.906 5  0.598  44.837 0.684 6 0.011(0.0086-0.014) 0.06 (0.043-0.098) 1.763 7 0.0029 (0.0025-0.0034) 0.0055(0.0046-0.0073) 4.708 8 0.0061 (0.005-0.0072) 0.015 (0.012-0.021) 3.2119 0.0033 (0.0027-0.004) 0.0087 (0.0069-0.012) 3.061 10 0.0051(0.0041-0.0062) 0.016 (0.013-0.023) 2.562 11 NE NE NE 12 0.023(0.017-0.03) 0.059 (0.042-0.116) 3.069 13 21.219 3713.114 0.571 150.0059 (0.0044-0.0076) 0.017 (0.012-0.029) 2.755 16 NE NE NE 17 0.02(0.015-0.026) 0.055 (0.04-0.092) 2.883 18 0.061 (0.045-0.094) 0.245(0.142-0.756) 2.131 NE = not effective at concentrations tested

TABLE 10 Response of X. cheopis adults after 24 h exposure. (95% CI inparentheses; LC₅₀ and LC₉₀ values expressed in terms of percentconcentration, wt:vol) Com- pound LC₅₀ LC₉₀ Slope 4 0.0059(0.0047-0.0075) 0.014 (0.011-0.023) 3.413 5 0.041 (0.034-0.049) 0.063(0.052-0.093) 6.925 6 0.017 (0.012-0.024) 0.075 (0.049-0.151) 2.022 70.0083 (0.0064-0.01) 0.019 (0.015-0.031) 3.527 8 0.0029 (0.002-0.0038)0.008 (0.0059-0.014) 2.949 9 0.0066 (0.0046-0.0086) 0.018 (0.013-0.032)2.9 10 0.0083 (0.0064-0.011) 0.021 (0.016-0.035) 3.143 11 NE NE NE 120.024 (0.018-0.034) 0.1 (0.065-0.196) 2.101 13 NE NE NE 15 0.0049(0.0039-0.0058) 0.0085 (0.0071-0.011) 5.366 16 NE NE NE 17 0.017(0.012-0.022) 0.059 (0.041-0.108) 2.326 18 0.064 (0.044-0.101) 0.484(0.284-1.609) 1.455 NE = not effective at concentrations tested

TABLE 11 Response of Ae. aegypti after 24 h exposure. (95% CI inparentheses; LC₅₀ and LC₉₀ values expressed in terms of percentconcentration, wt:vol) Com- pound LC₅₀ LC₉₀ Slope 4 0.0051 0.014 2.908 50.015 (0.008-0.029) 0.037 (0.031-0.162) 3.199 6 0.027 0.059 3.696 70.0057 0.0092 6.22 8 0.0046 (0.004-0.0053) 0.0087 (0.0072-0.011) 4.635 90.0075 0.021 2.845 10 0.0034 (0.0024-0.0045) 0.014 (0.01-0.023) 2.094 110.852 7.869 1.327 12 0.004 (0.0032-0.0048) 0.01 (0.008-0.014) 3.201 130.295 (0.21-0.522) 1.911 (0.922-7.257) 1.581 15 0.0024 0.0034 8.714 16NE NE NE 17 0.223 (0.158-0.402) 2.001 (0.873-10.537) 1.346 18 0.0590.114 4.472 NE = not effective at concentrations tested

TABLE 12 Residual activivy against Ixodes scapularis nymphs at 1, 2, and4 weeks. (95% CI in parentheses; LC₅₀ and LC₉₀ values expressed in termsof percent concentration, wt:vol) Compound Weeks LC₅₀ LC₉₀ Slope 6 12.062 524.656 0.533 6 2 NE NE NE 6 4 NE NE NE 7 1 0.023  0.13 1.682 7 20.026 (0.018–0.039) 0.168 (0.094–0.448) 1.586 7 4 0.026 (0.018–0.039)0.164 (0.093–0.429) 1.609 8 1 0.025  0.071 2.871 8 2 0.019 (0.014–0.024)0.056 (0.04–0.094) 2.727 8 4 0.25  0.71 2.871 9 1 0.0084 (0.005–0.013)0.08 (0.045–0.22) 1.304 9 2 0.0089 (0.0068–0.012) 0.027 (0.02–0.045)2.649 10 1 0.0071 (0.0055–0.0091) 0.019 (0.014–0.031) 3.005 10 2 0.0083(0.0057–0.011) 0.044 (0.029–0.081) 1.781 10 4 0.031  0.303 1.285 15 10.026 (0.02–0.033) 0.072 (0.052–0.121) 2.859 NE = not effective atconcentrations tested

TABLE 13 Residual activivy against Xenopsylla cheopis adults nymphs at1, 2, and 4 weeks. (95% CI in parentheses; LC₅₀ and LC₉₀ valuesexpressed in terms of percent concentration, wt:vol) Com- pound WeeksLC₅₀ LC₉₀ Slope 6 1 NE NE NE 7 1 0.016 (0.012–0.02) 0.05 (0.036–0.081)2.546 7 2 0.031 0.134 2.033 8 1 0.02 (0.016–0.026) 0.053 (0.04–0.082)3.107 8 2 0.018 (0.013–0.024) 0.081 (0.054–0.156) 1.931 8 4 0.035(0.026–0.051) 0.161 (0.098–0.387) 1.95  9 1 0.043 0.259 1.633 9 2 0.0270.085 2.571 9 4 0.43 (0.034–0.056) 0.111 (0.08–0.193) 3.135 10 1 0.03(0.022–0.041) 0.113 (0.074–0.226) 2.218 10 2 0.039 (0.029–0.053) 0.131(0.087–0.268) 2.442 15 1 0.059 0.148 3.211 NE = not effective atconcentrations tested

TABLE 14 Residual activivy against Aedes aegypti adults nymphs at 1, 2,and 4 weeks. (95% CI in parentheses; LC₅₀ and LC₉₀ values expressed interms of percent concentration, wt:vol) Compound Weeks LC₅₀ LC₉₀ Slope 61 0.132 (0.096–0.236) 0.659 (0.329–2.855) 1.833 6 2 NE NE NE 6 4 NE NENE 7 1 0.02 (0.014–0.029) 0.054 (0.041–0.105) 3.043 7 2 0.019(0.016–0.022) 0.046 (0.037–0.061) 3.342 7 4 0.013 (0.011–0.015) 0.038(0.031–0.051) 2.698 8 1 0.0065 0.014 3.858 8 2 0.0067 (0.0058–0.0076)0.012 (0.01–0.015) 4.953 8 4 0.02 0.29 7.772 9 1 0.0089 (0.0077–0.01)0.018 (0.015–0.023) 4.336 9 2 0.0042 (0.0036–0.0049) 0.0088(0.0074–0.011) 4.014 9 4 0.018 0.036 4.169 10 1 0.024 (0.02–0.028) 0.058(0.046–0.081) 3.286 10 2 0.014 (0.012–0.016) 0.021 (0.019–0.027) 6.93210 4 0.013 (0.0089–0.017) 0.04 (0.029–0.074) 2.645 15 1 0.011(0.0095–0.013) 0.027 (0.022–0.037) 3.392 15 2 0.014 (0.0074–0.025) 0.037(0.029–0.127) 3.136 NE = not effective at concentrations tested

Example 10 Pesticidally Acceptable Compositions

Compositions suitable for pesticidal uses are described, includingsolid, liquid, and gaseous formulations.

Dusts Component Amount (by weight) Dust A 13-hydroxy-valencene 2% highlydispersed silica 1% talcum 97% Dust B valencene-11,12-epoxide 1% highlydispersed silica 5% talcum 94% Dust C valencene-13-aldehyde 1%valencene-11,12-epoxide 1% sodium sulfate 98%Ready-to-use dusts may be obtained by intimately mixing the carrierswith the active ingredients.Emulsifiable Concentrate

Emulsifiable Concentrate Component Amount (by weight)nootkatone-1,10-epoxide 2% octylphenol polyethoxylate 3% calciumdodecylbenzenesulfonate 3% polyethoxylated castor oil 2% cyclohexanone35% xylene mixture 55%An emulsions of a desired concentration may be prepared from thisconcentrate by dilution with water.Extruder Granules

Extruder Granules Component Amount (by weight) 13-hydroxy-valencene 10%sodium ligninsulfonate 2% carboxymethyl cellulose 1% kaolin 87%The active ingredient is mixed with the additives, the mixture is groundtogether, water is added to the mixture, and the mixture is thenextruded, granulated, and subsequently dried.

Example 11 Production of Compound 15 (Valencene-13-aldehyde)

Valencene-13-aldehyde was produced by the reaction:

using the following procedure: one gram (4.89 mmol) of valencene wasdissolved in 10 ml of dry pyridine. The solution was stirred and 1 g(9.01 mmol) of SeO₂ was added to the reaction. The mixture was refluxedfor 5 hours until the yellow solution turned black. The mixture wasfiltered to eliminate the selenium dust. The brown solution was passedthrough Silica gel-Na₂CO₃ 1:1 and the funnel was washed with ether. Thepyridine was removed by vacuum distilation and the remaining oil wasanalyzed by chromatography through Silica gel-Na₂CO₃ 1:1 with hexane aseluent recovering the most polar fraction (Rf =0.2 in hexane). Aftersolvent evaporation, a yellow oil (0.125 g, 11.59% yield) was obtained,showing just one product in high purity.

Example 12 Production of Compound 16 (Nootkatone-11,12-epoxide)

Nootkatone-11,12-epoxide was produced by the reaction:

using the following procedure: four grams of nootkatone (18.32 mmol)were dissolved in 30 ml of diethyl ether and 3.79 g (18.32 mmol) ofm-chloroperbenzoic acid (mCPBA 80%) were added while stirring. After twohours, an excess of one mol (3.79 g) of mCPBA 80% was added. Thesolution was stirred for two hours more, then 30 ml of cold water and 30ml of NaHCO₃ saturated solution was added to stop the reaction. Themixture was poured through a separatory funnel to separate the organiclayer. The remaining water layer was washed twice with diethyl ether (30ml) and joined with the first one. The organic layer containing diethylether and the product was dried with anhydrous sodium sulfate, and etherwas removed with rotavap or by nitrogen flow yielding 2.5 g (58% yield)of product as a coalescent pale yellow oil that eventually crystallized(rf=0.24 in hexane-diethyl ether 1:1).

The principle epoxide product showed a high purity by NMR analysis(>90%). However, some efforts to purify the compound by silicagel-sodium carbonate chromatography yielded a mixture of open products,likely diols. Some crystals of the pure product were obtained by passinga small quantity of the crude epoxide through a Na₂CO₃-silica gel (7:3)column yielding white crystals that melted at 35.2-35.7° C.

Example 13 Production of Compound 17 (Nootkatone-1,10-epoxide)

Nootkatone-1,10-epoxide was produced by the reaction:

using the following procedure: five grams (22.29 mmol) of nootkatonewere dissolved in 30 ml of methanol, the solution was cooled to 10° C.while stirring, then 4.67 g (133.74 mmol, 15.56 ml) of H₂O₂ (30%) wasadded. When the addition was finished, 10 ml of KOH 6N was added drop bydrop over a period of 20 minutes, taking care that the temperature didnot exceed 10° C. After the KOH addition, the mixture was stirred for 3hours at 25° C., the methanol was evaporated in a rotavap, and theproduct was extracted from the water solution by diethyl ether (3×20ml). The organic solution with the product was dried on anhydrous Na₂SO₄and the ether completely eliminated by evaporation to give 2.97 g (44.4%yield) of a colorless oil (rf=0.44 in Hexane-Acetone 9:1), NMR analysisof the product showed high purity.

Example 14 Production of Compound 18 (Nootkatone-1,10-11,12-diepoxide)

Nootkatone-1,10-11,12-diepoxide was produced by the reaction:

using the following procedure: 1.7 g (7.26 mmol) of the epoxide weredissolved in 50 ml of anhydrous methanol, the solution was cooled to 10°C., then 2.5 ml (0.75 g, 22.06 mmol) was added drop by drop whilestirring, over a period of 20 minutes, taking care that the temperaturedid exceed 10° C. After the KOH addition, the mixture was stirred for 3hours then killed with 30 ml of cold water. The methanol was eliminatedin a rotavap and the water solution was extracted with diethyl ether(3×20 ml). The ethereal solution containing the product was evaporatedby rotavap or by nitrogen flow, yielding 0.735 g (40.5%) of thediepoxide as a semisolid white, (rf=0.50 in hexane-acetone 1:1). ¹³C-NMRshowed a mixture of diastereo isomers with high purity.

Having illustrated and described the principles of the invention byseveral embodiments, it should be apparent that those embodiments can bemodified in arrangement and detail without departing from the principlesof the invention. Thus, the invention as claimed includes all suchembodiments and variations thereof, and their equivalence, as comewithin the true spirit and scope of the claims stated below.

1. A method for controlling an arthropod, comprising contacting anarthropod selected from the taxonomic order or subclass Acarina,Anoplura, Blattaria, Homoptera, Hymenoptera, Lepidoptera, orSiphonaptera, with a pesticidally effective amount of a pest controlagent comprising isolated nootkatone, wherein the nootkatone is presentin a concentration of at least about 0.01 ppm (parts per million) andless than 65×10⁻³ percent (wt:vol), and a pesticidally acceptablecarrier.
 2. The method of claim 1, wherein controlling the arthropodcomprises killing the arthropod.
 3. The method of claim 1, whereincontrolling the arthropod comprises repelling the arthropod.
 4. Themethod of claim 1, wherein the pest control agent is applied directly tothe arthropod.
 5. The method of claim 1, wherein the pest control agentis applied to a locus comprising the arthropod.
 6. The method of claim1, wherein the method comprises an area-wide application of the pestcontrol agent.
 7. The method of claim 1, wherein the pest control agentis provided to a human or non-human animal.
 8. The method of claim 7,wherein the pest control agent is orally administered or provided as atopical treatment.
 9. The method of claim 1, wherein the pest controlagent is embedded within a material.
 10. The method of claim 9, whereinthe material is a collar, ear tag, piece of clothing, siding, wallstuds, or beam.
 11. The method of claim 1, wherein the pest controlagent is applied to plants, animals or objects within an environmentthat comes into contact with the arthropod.
 12. The method of claim 1,wherein the arthropod is a member of the taxonomic order Siphonaptera.13. The method of claim 12, wherein the arthropod is Xenopsylla cheopis(rat flea).
 14. The method of claim 1, wherein the arthropod is a memberof the Ixodes species.
 15. The method of claim 14, wherein the arthropodis Ixodes scapularis (deer tick).
 16. The method of claim 1, furthercomprising identifying the arthropod as a vector for a disease.
 17. Themethod of claim 16, wherein the disease is Lyme disease; tickborne-babesiosis; tularemia; powassan-like virus infection; tick borneencephalitis; relapsing fever; Colorado Tick Fever; ehrlichiosis; RockyMountain Spotted Fever; or the Plague.
 18. The method of claim 1,wherein the isolated nootkatone is present in a concentration of atleast about 0.1 ppm (parts per million).
 19. The method of claim 1,wherein the nootkatone comprises synthetic nootkatone.
 20. The method ofclaim 1, wherein the nootkatone is isolated from Alaska yellow cedar.21. A method for controlling an arthropod, comprising contacting anarthropod selected from the taxonomic order or subclass Acarina,Anoplura, Blattaria, Homoptera, Hymenoptera, Lepidoptera, orSiphonaptera with a pesticidally effective amount of a pesticidalcomposition consisting essentially of nootkatone and a pesticidallyacceptable carrier, wherein the nootkatone is present in a concentrationof at least about 0.01 ppm (parts per million) and less than 65×10⁻³percent (wt:vol).
 22. The method of claim 21, wherein controlling thearthropod comprises killing the arthropod.
 23. The method of claim 21,wherein controlling the arthropod comprises repelling the arthropod. 24.A method for controlling an arthropod, comprising contacting anarthropod selected from the taxonomic order or subclass Acarina,Anoplura, Blattaria, Homoptera, Hymenoptera, Lepidoptera, orSiphonaptera with a pesticidally effective amount of a pesticidalcomposition comprising a pesticidal eremophilane sesquiterpene and apesticidally acceptable carrier; wherein the pesticidal eremophilanesesquiterpene consists of nootkatone, and wherein the nootkatone ispresent in a concentration of at least about 0.01 ppm (parts permillion) and less than 65×10⁻³ percent (wt:vol).
 25. The method of claim1, wherein the arthropod is a member of the taxonomic order Anoplura.26. The method of claim 25, wherein the arthropod is a member of thePediculus species.
 27. The method of claim 21, wherein the isolatednootkatone is present in a concentration of at least about 0.1 ppm(parts per million).